| genetic study of suppressor-sensitive mutants of the bacillus subtilis bacteriophage phi 29. | with bacteriophage phi29 of bacillus subtilis 133, suppressor-sensitive (sus) hydroxylamine mutants have been isolated. intracistronic and intercistronic quantitative complementation placed the mutants in 13 cistrons, and three-factor crosses have been used to assign an unambiguous order for 10 cistrons. recombination frequencies have been presented for several regions of the genome to facilitate comparison of the sus system with the previously published temperature-sensitive mapping systems. | 1973 | 4196635 |
| synthesis of bacteriophage phi 29 proteins in bacillus subtilis. | seventeen bacteriophage phi29 proteins were detected in ultraviolet light-irradiated bacillus subtilis by autoradiography of polyacrylamide slab gels. the appearance of phi29 proteins occurred either before or concomitantly with viral dna replication. viral proteins detected early in the infectious cycle consisted of nine polypeptides ranging from 5,200 daltons to 54,000 daltons. two of the early proteins were identified as, respectively, the major capsid protein and the protein comprising the f ... | 1973 | 4199108 |
| proteins induced in bacillus subtilis infected with bacteriophage phi 29. | | 1973 | 4200881 |
| antigenic properties of bacteriophage phi 29 structural proteins. | serological methods and electron microscopy were used to study the structural proteins of the small bacillus subtilis bacteriophage phi29. this virus has a large number of fibers attached at both ends of its prolate head. a complex neck assembly is comprised of 12 symmetrically arranged appendages as the outer component. head fibers, neck appendages, and the head surface bind anti-phi29 antibodies. immune sera absorbed with defective lysates of suppressor-sensitive (sus) mutants have been used t ... | 1973 | 4202619 |
| viral protein synthesis in bacteriophage phi 29-infected bacillus subtilis. | twenty-three (14)c-labeled phage phi29-specific proteins in lysates of uv-irradiated bacillus subtilis have been resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis and identified by autoradiography. included in this group of proteins are the six major structural proteins of the virion. analysis of the temporal sequence of viral protein synthesis indicates that three groups of proteins can be identified by time of appearance, beginning at 2 to 4, 4 to 6, or 8 to 10 min after in ... | 1973 | 4203085 |
| analysis of bacteriophage phi 29 gene function: protein synthesis in suppressor-sensitive mutant infection of bacillus subtilis. | phage phi29 suppressor-sensitive (sus) mutants of 14 cistrons have been examined for production of (14)c-labeled viral-specific proteins in restrictive infections of bacillus subtilis. proteins specified by four cistrons (h, j, l, and n) have been resolved and identified by sodium dodecyl sulfate gel electrophoresis and autoradiography, and fragments of the normal polypeptides were detected. mutants of six cistrons (c, d, e, f, i, and m) demonstrated two or more missing bands in the gel profiles ... | 1974 | 4204249 |
| a precursor of the neck appendage protein of b. subtilis phage phi 29. | | 1974 | 4212893 |
| suppressor-sensitive mutants and genetic map of bacillus subtilis bacteriophage phi 29. | | 1974 | 4214301 |
| gene expression during the development of bacteriophage phi 29. 3. analysis of viral-specific protein synthesis with suppressible mutants. | fifty-four suppressible mutants of bacteriophage phi29 have been isolated with a variety of mutagens and assigned to eight complementation groups. viral-specific protein synthesis in uv light-irradiated, nonsuppressing bacillus subtilis 60084 was analyzed with exponential acrylamide gels. four additional phi29 proteins which were undetected on ordinary acrylamide gels are reported in this paper. five phage phi29 proteins have been unambiguously assigned to specific cistrons. two cistrons had ple ... | 1974 | 4362871 |
| characterization of the phage phi 29 protein p5 as a single-stranded dna binding protein. function in phi 29 dna-protein p3 replication. | the phage phi 29 protein p5, required in vivo in the elongation step of phi 29 dna replication, was highly purified from escherichia coli cells harbouring a gene 5-containing plasmid and from phi 29-infected bacillus subtilis. the protein was characterized as the gene 5 product by amino acid analysis and nh2-terminal sequence determination. the purified protein p5 was shown to bind to single-stranded dna and to protect it against nuclease degradation. no effect of protein p5 was observed either ... | 1989 | 2499869 |
| transcription during the development of bacteriophage phi 29: production of host- and phi 29-specific ribonucleic acid. | the synthesis of ribonucleic acid (rna) during development of the virulent bacillus subtilis bacteriophage phi29 has been analyzed. transcription of host deoxyribonucleic acid (dna) continues at the preinfection rate throughout the latent period of viral growth. rna-dna hybridization was used to show that host messenger rna synthesis continues late into the phage lytic cycle. amino acid-labeling experiments show that this rna is continuously used to produce protein. ribosomal rna production is n ... | 1972 | 4630153 |
| regions at the carboxyl end of bacteriophage phi 29 protein p6 required for dna binding and activity in phi 29 dna replication. | series of deletions corresponding to the carboxyl end of the phage phi 29 protein p6 have been constructed and their activity in the initiation of phi 29 dna replication and their capacity to interact with the phi 29 dna ends have been studied. determination of the activity of the deletion mutants in phi 29 dna replication indicated the dispensability of the 14 carboxy-terminal amino acids of the protein. the activity of protein p6 decreased with deletions from 23 to 39 amino acids and was undet ... | 1989 | 2501757 |
| structure of bacillus subtilis bacteriophage phi 29 and the length of phi 29 deoxyribonucleic acid. | anderson, d. l. (university of minnesota, minneapolis), d. d. hickman, and b. e. reilly. structure of bacillus subtilis bacteriophage phi29 and the length of phi29 deoxyribonucleic acid. j. bacteriol. 91:2081-2089. 1966-bacillus subtilis bacteriophage phi29 were negatively stained with phosphotungstic acid. the head of phi29 has a hexagonal outline with a flattened base, and is about 315 a wide and 415 a in length. the virus has an intricate tail about 325 a in length. twelve spindle-shaped appe ... | 1966 | 4957028 |
| physical and biological properties of phage phi 29 deoxyribonucleic acid. | deoxyribonucleic acid (dna) molecules having a mean length of 5.8 mum were released from purified bacillus subtilis bacteriophage phi29 with 2 m sodium perchlorate. small 0.1 to 0.2-mum molecules were also detected in these dna preparations. since intact single chains annealed to form linear duplex molecules, phage phi29 dna was found to be nonpermuted. the molecular weights of single chains of phi29 dna were approximately half that of native dna, as determined by analytical band sedimentation i ... | 1968 | 4972802 |
| rescue of genetic markers from bacteriophage phi 29 dna fragments. | | 1970 | 4998256 |
| a genetic study of temperature-sensitive mutants of the bacillus subtilis bacteriophage phi 29. | | 1971 | 5000908 |
| dna-protein complex in circular dna from phage phi-29. | | 1971 | 5002464 |
| mapping of temperature sensitive mutants of bacteriophage phi 29. | | 1972 | 5018452 |
| characterization of a new prokaryotic transcriptional activator and its dna recognition site. | the expression of the bacillus subtilis phage phi 29 dna is controlled by the viral gene 4 product, which is required for the initiation of transcription at the unique late promoter a3. protein p4 binds specifically to a phi 29 dna fragment containing the a3 promoter. dnase i footprinting analysis has shown that the dna binding region for protein p4 is located between nucleotides -50 and -100 relative to the transcription start site. methylation interference assays suggest that two eight base-pa ... | 1989 | 2504924 |
| temperature-sensitive mutants of bacteriophage phi-29. | | 1971 | 5167656 |
| effect of elevated temperature on deoxyribonucleic acid synthesis in bacteriophage phi-29-infected bacillus amyloliquefaciens. | deoxyribonucleic acid (dna) synthesis in bacteriophage phi29-infected bacillus amyloliquefaciens was studied at 37 and 45 c. infectious intracellular particles appear at the same time at both temperatures, but the average burst size is reduced 45 to 50% at 45 c. there is a transient inhibition of cellular mass increase at 45 c which is not observed at the lower temperature. in addition, the rate of host dna synthesis is reduced and the onset of viral-specific dna replication is delayed for 6 to ... | 1970 | 5497888 |
| cloning and expression of gene 2, required for the protein-primed initiation of the bacillus subtilis phage phi 29 dna replication. | a phi 29 dna fragment containing gene 2, coding for a phi 29-specific dna polymerase required for the formation of the terminal protein p3-damp initiation complex, the first step in phi 29 dna replication, has been cloned in plasmid pplc28 under the control of the pl promoter of bacteriophage lambda. four polypeptides of mr 68 000, 5800 and 3400 and less than 2000 were labelled with [35s]methionine after heat induction. the protein of mr 68 000 had the size expected for protein p2 and it account ... | 1984 | 6092229 |
| cloning, nucleotide sequence and high level expression of the gene coding for the connector protein of bacillus subtilis phage phi 29. | the phi 29 dna restriction fragment hindiii-d, shown to contain gene 10 coding for the connector protein, has been cloned in plasmid pplc28 under the control of the pl promoter of phage lambda. after heat induction to inactivate the lambda repressor, a protein with the electrophoretic mobility of the connector protein p10 was synthesized, accounting for about 30% of the total escherichia coli protein after 3 h of induction. the 2205 nucleotide-long sequence of the cloned hindiii-d fragment has b ... | 1984 | 6096227 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: oriented and quantized in vitro packaging of dna protein gp3. | the assembly of phage phi 29 occurs by a single pathway, and the dna protein (dna-gp3) of "packaging intermediates" can be obtained after dnase i interruption of in vitro complementation. a broad spectrum of dna molecules of variable length was isolated from dnase i-treated proheads. restriction endonuclease ecori digestion and electrophoretic analysis of these dna molecules suggested that dna-gp3 packaging was oriented with respect to the physical map and was a complex process. proteinase k-tre ... | 1983 | 6185695 |
| effects of internal deletions on the priming activity of the phage phi 29 terminal protein. | a series of internal deletions of gene 3, coding for the phage phi 29 dna terminal protein, have been constructed and characterized. in addition, a substitution mutant in the sequence corresponding to amino acids (aa) 49-51 was obtained. the priming activity of the substitution mutant protein, in the formation of the protein p3-damp initiation complex, was drastically reduced suggesting that some of the aa present at position 49-51 are essential for p3 function. deletions of 8 to 33 aa, from aa ... | 1989 | 2511080 |
| in vitro packaging of bacteriophage phi 29 dna restriction fragments and the role of the terminal protein gp3. | restriction fragments of bacteriophage phi 29 dna-gp3 (dna-gene product 3 complex) were packaged in a completely defined in vitro system that included purified proheads, the dna packaging protein gp16 and atp. both left and right end dna-gp3 fragments were packaged in this system, in contrast to the oriented and selective packaging of left end dna-gp3 fragments in extracts; left ends could be packaged quantitatively in the defined system, while the packaging efficiency of right ends was generall ... | 1989 | 2530357 |
| signals at the bacteriophage phi 29 dna replication origins required for protein p6 binding and activity. | protein p6 of bacillus subtilis phage phi 29 binds specifically to the ends of the viral dna that contain the replication origins, giving rise to a nucleoprotein structure. dna regions recognized by protein p6 have been mapped by deletion analysis and dnase i footprinting. main protein p6-recognition signals have been located between nucleotides 62 and 125 at the right phi 29 dna end and between nucleotides 46 and 68 at the left end. in addition, recognition signals are also present at other sit ... | 1989 | 2767056 |
| high level synthesis in escherichia coli of the bacillus subtilis phage phi 29 proteins p3 and p4 under the control of phage lambda pl promoter. | the hind iii g fragment from the bacillus subtilis phage phi 29 dna, inserted downstream from the bacteriophage lambda promoter pl carried by a pbr322 derivative plasmid (pplc28), directed the synthesis in e. coli of two proteins of apparent molecular weight 27500 and 12500. with the use of the recombinants obtained with the dna from mutants sus3(91) and sus4(56), the two proteins were identified as a modified p3 (p3'), the protein covalently linked to the 5' ends of phi 29 dna, and p4, responsi ... | 1982 | 6292851 |
| in vivo transcription of bacteriophage phi 29 dna: transcription initiation sites. | the initiation sites of the rna transcripts synthesized in vivo in bacillus subtilis infected with bacteriophage phi 29 have been mapped by s1 protection experiments. nine transcription initiation sites were localized along the entire phi 29 genome, close to previously reported b. subtilis and escherichia coli rna polymerase-binding sites. eight of these sites corresponded to early transcription and only one corresponded to late transcription. by using 5'-end-labeled rna, four of the early sites ... | 1986 | 3023677 |
| cloning and expression in escherichia coli of the gene coding for the protein linked to the ends of bacillus subtilis phage phi 29 dna. | a phi 29 dna fragment containing gene 3, coding for the 5'-terminal protein, and several other early genes has been cloned in a pbr322 derivative plasmid (pkc30) under the control of the pl promoter of bacteriophage lambda. four polypeptides of mr 27000, 18500, 17500 and 12500 were labelled with [35s]methionine after heat induction, accounting for about 15% of the de novo synthesized protein. the mr 27000 and 12500 proteins were characterized as p3, the 5'-terminal protein, and p4, involved in t ... | 1983 | 6301951 |
| related functional domains in virus dna polymerases. | analysis of the lesions in several drug-resistant dna polymerase mutants of herpes simplex virus along with comparative analysis of the published polymerase sequences of other human herpesviruses has shown that most lesions (five out of six) are substitutions at amino acid residues conserved in all four polymerases. furthermore, the majority of lesions are in regions of the polypeptide where there are marked clusterings of conserved residues. on the basis of these data we have identified several ... | 1987 | 3034575 |
| initiation of phage phi 29 dna replication by the terminal protein modified at the carboxyl end. | a mutant at the carboxyl end of the terminal protein, p3, of phage phi 29 dna has been constructed by inserting an containing the stop translation codon tga in the three possible reading frames, immediately downstream of a phage phi 29 dna fragment coding for all but the last five amino acids of protein p3. the activity in the formation of the p3-damp initiation complex in vitro of this mutant as well as another one previously isolated, also mutated at the carboxyl end, have been tested. the re ... | 1983 | 6316260 |
| template requirements for initiation of phage phi 29 dna replication in vitro. | the template requirements for the formation of the phi 29 protein p3-damp initiation complex in vitro have been studied. the initiation reaction requires the parental protein p3 but not an intact dna molecule. protein p3-containing fragments from the left- or right-hand dna ends were active as template for formation of the initiation complex provided they had a minimal size: a 26-base-pair-long fragment was active whereas a 10-base-pair-long one was essentially inactive. however, the activity of ... | 1984 | 6320176 |
| in vitro transcription of the bacillus subtilis phage phi 29 dna by bacillus subtilis and escherichia coli rna polymerases. | the escherichia coli rna polymerase bound to phage phi 29 dna has been visualized by electron microscopy. thirteen specific binding sites have been observed at 1.7,2.6,5.5,10.4,13.7,25.2,25.7,26.3,33.5,59.5,69.2,91.7 and 99.6 dna length units and they have been named a1,a1i,a1ii,a1iii,a1iv,a2,a2i, a3, a4,b1,b1i,c1 and c2, respectively. the binding sites a1,a2,a3,b1,c1 and c2 coincide with those found with bacillus subtilis rna polymerase. the transcription of phage phi 29 dna with b. subtilis or ... | 1984 | 6322128 |
| overproduction and purification of the connector protein of bacillus subtilis phage phi 29. | a phi 29 dna fragment containing genes 10 and 11, coding for the connector protein and the lower collar protein, respectively, has been cloned in the pbr322 derivative plasmid pkc30 under the control of the pl promoter of phage lambda. two polypeptides with the electrophoretic mobility of proteins p10 and p11 were labelled with 35s-methionine after heat induction. the proteins were characterized as p10 and p11 by radioimmunoassay and they represented about 10% and 7%, respectively, of the total ... | 1984 | 6324116 |
| restoration of direct fourier three-dimensional reconstructions of crystalline specimens by the method of convex projections. | we consider the problem of the three-dimensional (3-d) reconstruction of objects by the direct fourier method (dfm) and their restoration by the method of projections on to convex sets (pocs). the main discussion is centered on the case of specimens arranged in a two-dimensional (2-d) crystal and imaged by transmission electron microscopy, although the conclusions could be extended to more general cases. we present results of the restoration of the 3-d reconstruction of a computer generated 2-d ... | 1987 | 3573036 |
| three-dimensional reconstruction of the connector of bacteriophage phi 29 at 1.8 nm resolution. | the three-dimensional reconstruction of the connector of bacteriophage phi 29 has been obtained from tilt series of negatively stained tetragonal ordered aggregates under low-dose conditions and up to a resolution of (1/1.8) nm-1. these connectors are built up as dodecamers of only one structural polypeptide (p10). two connectors form the crystal unit cell, each one facing in the opposite direction with respect to the plane of the crystal and partially overlapping. the main features of the two c ... | 1986 | 3586012 |
| nucleotide sequence of phage phi 29 gene 7: structure of intergenic spacer between the major early and late genes. | | 1986 | 3763399 |
| factors involved in the initiation of phage phi 29 dna replication in vitro: requirement of the gene 2 product for the formation of the protein p3-damp complex. | to study the requirements for the in vitro formation of the protein p3-damp complex, the first step in phi29 dna replication, extracts from b. subtilis infected with phi29 mutants in genes 2, 3, 5, 6 and 17, involved in dna synthesis, have been used. the formation of the initiation complex is completely dependent on the presence of a functional gene 2 product, in addition to protein p3 and phi29 dna-protein p3 as template. atp is also required, although it can be replaced by other nucleotides. t ... | 1983 | 6402761 |
| assembly of the tail protein of the bacillus subtilis phage phi 29. | by in vitro complementation we have determined that gene 13 product functions during phi 29 morphogenesis after the formation of 11- particles, specifically in the functional assembly of the tail protein, p9. protein p9 from 8- but not from 8-13- extracts assembles in vitro into either 11-13- or 12-13- particles. the action of gene 13 product on p9 for its correct assembly has to take place in vivo; no complementation of 12-13- and 9- lysates occurs in vitro. protein p9, isolated from phi 29-inf ... | 1983 | 6402855 |
| replication of phage phi 29 dna with purified terminal protein and dna polymerase: synthesis of full-length phi 29 dna. | a system that replicates bacteriophage phi 29 dna with protein p3 covalently attached to the two 5' ends, using as the only proteins the phi 29 dna polymerase and the terminal protein, is described. restriction analysis of the 32p-labeled dna synthesized in vitro showed that all phi 29 dna fragments were labeled. analysis by alkaline sucrose gradient centrifugation of the dna labeled during a 10-min pulse showed that, after a 20-min chase, about half of the dna molecules had reached apparently f ... | 1985 | 3863101 |
| protein-primed initiation of phage phi 29 dna replication. | we recently reported the development of an in vitro replication system for bacteriophage phi 29 dna. we have used this system for the isolation of replication activity associated with gene 3 protein (terminal protein) from phi 29-infected bacillus subtilis cells. we utilized two assay systems: (i) dna replication dependent on phi 29 dna with the 5' end covalently linked to terminal protein (dna-protein) and (ii) the formation of complex between the terminal protein and damp. the dna-replication ... | 1983 | 6410387 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: prohead restoration for dna-gp3 packaging and assembly. | the dna-protein complex dna-gp3 of phi 29 is efficiently packaged into purified proheads with the aid of plasmid-derived gp16. the filled heads can be assembled to phage by addition of an extract providing the products for neck-tail assembly (bjornsti et al., j. virol. 50:766-772, 1984). however, purified proheads lost their competence to package dna-gp3 upon storage for 2 months at 4 degrees c. competence was restored by complementation with extracts of certain mutant-infected cells, and these ... | 1985 | 3919187 |
| inefficient translation of t7 late mrna by bacillus subtilis ribosomes. implications for species-specific translation. | bacillus subtilis 30 s subunits inefficiently recognize initiation sites in mrnas from gram-negative bacteria, but they are able to efficiently recognize initiation sites in mrna derived from gram-positive bacteria. mclaughlin et al. (mclaughlin, j. r., murray, c. l., and rabinowitz, j. c. (1981) j. biol. chem. 256, 11283-11291) have suggested that b. subtilis ribosomes require a strong shine-dalgarno sequence for translation initiation. to test whether this criterion is sufficient to explain th ... | 1985 | 3934154 |
| structure of phage phi 29 connector protein assembled in vivo. | the protein p10 that forms the connector of phage phi 29, has been produced in escherichia coli harboring a plasmid that carried the gene coding for this protein. the connector protein is assembled in a 13.4-s oligomer that has an apparent molecular weight of 460,000, suggesting that it is a dodecamer. the purified oligomers have been studied by electron microscopy of the isolated particles as well as by image-processing techniques (fourier and rotational filtering) of artificially induced two-d ... | 1985 | 3936270 |
| initiation events in in-vitro packaging of bacteriophage phi 29 dna-gp3. | initiation events in the packaging of bacteriophage phi 29 dna-gp3 (dna-gene product 3 complex) were studied in a completely defined in-vitro system that included purified proheads, dna-gp3 and the dna packaging protein gp16. in the sequential interactions, gp16 first bound to, and was modified by, the prohead. the prohead-gp16 complex then bound to dna-gp3, resulting in a second modification of gp16 that permitted binding of atp. dna-gp3 aggregates were produced, and the hydrolysis of atp accom ... | 1987 | 3119862 |
| nucleotide sequence and transcription of a bacteriophage 29 early promoter. | we have studied the in vitro and in vivo transcription of a promoter for bacillus subtilis rna polymerase on bacteriophage phi 29 dna. the promoter is identified as an early promoter as it is transcribed in vitro by uninfected b. subtilis sigma 55-containing rna polymerase; is transcribed in vivo at both 7 min after infection and in the presence of chloramphenicol; and is transcribed right to left on the standard phi 29 map. the nucleotide sequence of the promoter and the initiation site for rna ... | 1985 | 3997806 |
| three-dimensional reconstruction of bacteriophage phi 29 neck particles at 2 x 2 nm resolution. | the three-dimensional structure of the head-to-tail connecting region of bacteriophage phi 29 has been studied by analysing two-dimensional, hexagonal ordered aggregates of negatively stained viral necks to a resolution of 2 x 2 nm. these necks are composed of two proteins, p10 and p11; p10 being the connector protein. a 12-folded and a 6-folded axially symmetric domain are present in the specimen. the 12-folded domain is the larger part of the structure; it consists of 12 subunits associated in ... | 1985 | 4009722 |
| bacteriophage phi 29 dna replication in vitro: participation of the terminal protein and the gene 2 product in elongation. | from phi 29-infected bacillus subtilis cells, we have isolated a protein fraction which promotes in vitro replication of phi 29 dna. this fraction catalyses both initiation and elongation, indicating that it contains the product of gene 3 (tp: terminal protein) and the product of gene 2 (gp2: probably a dna polymerase), since initiation requires the two products (blanco et al. 1983; matsumoto et al. 1983). the fractions isolated from cells infected with temperature-sensitive (ts) mutants of gene ... | 1984 | 6438445 |
| transfecting deoxyribonucleic acid of bacillus bacteriophage phi 29 that is protease sensitive. | the transfecting activity of bacillus phage varphi29 dna, extracted either by sodium lauroyl sarcosine-phenol or by 2 m perchlorate, was destroyed by treatment with proteolytic enzymes, although these enzymes did not effect transfecting dnas of spp1, spo1, and sp50. these facts suggest that a protein is associated with transfective varphi29 dna. stabilization of protease-resistance during transfection appeared earlier than that of dnaseresistance, indicating that the protein associated with varp ... | 1972 | 4504368 |
| structure of replicating dna molecules of bacillus subtilis bacteriophage phi 29. | we isolated phi 29 dna replicative intermediates from extracts of phage-infected bacillus subtilis, pulsed-labeled with [3h]thymidine, by velocity sedimentation in neutral sucrose followed by cscl equilibrium density gradient centrifugation. during a chase, the dna with a higher sedimentation coefficient in neutral sucrose and a lower sedimentation rate in alkaline sucrose than that of viral phi 29 dna was converted into mature dna. the material with a density higher than that of mature phi 29 d ... | 1980 | 6768899 |
| the protein covalently linked to the 5' termini of the dna of bacillus subtilis phage phi 29 is involved in the initiation of dna replication. | | 1980 | 6771916 |
| gene expression during the development of bacillus subtilis bacteriophage phi 29. i. analysis of viral-specific transcription by deoxyribonucleic acid-ribonucleic acid competition hybridization. | the ribonucleic acid (rna) specified by bacteriophage phi29 was analyzed to determine its composition at various times in the viral lytic cycle. although viral-specific rna was detected immediately after infection, a large increase in the rate was observed at 10 min when dna synthesis began. phi29 was found to resemble other viruses in that gene expression occurred in two stages which could be defined temporally as "early" and "late." early rna appeared before the onset of viral deoxyribonucleic ... | 1973 | 4630802 |
| temperature-shift analysis of bacteriophage phi 29 gene expression in bacillus amyloliquefaciens. | temperature shift-up experiments with conditional lethal mutants of bacillus phage phi29 have allowed placement of early, middle, and late functions on the linear phi29 genetic map most of the phi29 cistrons are late and are found at the ends of the map. | 1971 | 5119489 |
| specificity of promoter site utilization in vitro by bacterial rna polymerases on bacillus phage phi 29 dna. transcription mapping with exonuclease iii. | bacillus subtilis rna polymerase holoenzyme transcribes phi 29 dna in vitro producing five major rna species defined by characteristic electrophoretic mobilities. in addition to these products, escherichia coli rna polymerase transcribes phi 29 dna to yield three rna species not detected when transcribing with the b. subtilis enzyme under the same optimal reaction conditions for rna synthesis. transcriptional analysis of purified restriction fragments and exonuclease iii-digested dna established ... | 1980 | 6251067 |
| interaction of the bacteriophage phi 29 protein p6 with double-stranded dna. | the bacillus subtilis bacteriophage phi 29 protein p6 binds to double-stranded dna, but not to single-stranded dna, as determined by a gel retardation assay. the nature of the interaction was further studied by dnase i "footprinting" experiments. protein p6 binds to fragments containing the right or left terminal sequences of phi 29 dna, producing a characteristic pattern of hypersensitive bands spaced about 24 nucleotides apart along most of the fragment, flanking protected regions. binding of ... | 1988 | 3124105 |
| nucleotide sequence at the termini of the dna of bacillus subtilis phage phi 29. | phage phi 29 dna cannot be phosphorylated with polynucleotide kinase and [gamma-32p]atp because of the presence of a viral protein covalently linked to the 5' termini. the 5' ends can, however, be made susceptible to phosphorylation by treatment with alkali and alkaline phosphatase. restriction fragments hpa ii c and hpa ii f, corresponding to the right and left ends of phi 29 dna, respectively, were labeled at the 5' ends with polynucleotide kinase and [gamma-32p]atp or at the 3' ends with term ... | 1981 | 6262800 |
| structure of protein-containing replicative intermediates of bacillus subtilis phage phi 29 dna. | | 1982 | 6801848 |
| nucleotide sequences of transcription and translation initiation regions in bacillus phage phi 29 early genes. | phi 29 dna directs the synthesis of three major proteins of mr = 22,400, 13,900, and 10,500 in a cell-free transcription-translation system derived from bacillus subtilis. we have determined the locations of the coding regions for these early proteins on the phi 29 genome, and our results are in agreement with genetic evidence that the 22.4-kilodalton protein is the product of cistron 17 (p17) and the 13.9-kilodalton protein is the product of cistron 6 (p6). the 13.9-kilodalton and 10.5-kilodalt ... | 1982 | 6274853 |
| morphogenesis of bacteriophage phi 29 of bacillus subtilis: dna-gp3 intermediate in in vivo and in vitro assembly. | the assembly of phage phi 29 occurs by a single pathway, and dna-protein (dna-gp3) has been shown to be an intermediate on the assembly pathway by a highly efficient in vitro complementation. at 30 degrees c, about one-half of the viral dna synthesized was assembled into mature phage, and the absolute plating efficiency of phi 29 approached unity. dna packaging at 45 degrees c was comparable to that at 30 degrees c, but the burst size was reduced by one-third. when cells infected with mutant ts3 ... | 1982 | 6804642 |
| rna polymerase of myxococcus xanthus: purification and selective transcription in vitro with bacteriophage templates. | dna-dependent rna polymerase from vegetative cells of the gram-negative, fruiting bacterium myxococcus xanthus was purified more than 300-fold by a modified burgess procedure (lowe et al., biochemistry 18:1344-1352, 1979), using polymin p precipitation, 40 to 65% saturated ammonium sulfate fractional precipitation, double-stranded dna cellulose chromatography, a5m gel filtration chromatography, and single-stranded dna agarose chromatography. the last step separated the rna polymerase into a core ... | 1982 | 6806251 |
| effect of the delta subunit of bacillus subtilis rna polymerase on initiation of rna synthesis at two bacteriophage phi 29 promoters. | initiation of rna synthesis by bacillus subtilis rna polymerase (sigma-43) has been examined at two early promoters of phage phi 29: the a2 promoter, which is a weak promoter, and the g2 promoter, which is a strong promoter. the delta subunit of the polymerase inhibits the rate of initiation at a2, but not g2. in addition, formation of stable complexes by the polymerase at a2, but not at g2, requires the presence of the first two nucleotides of the a2 transcript. | 1987 | 3126800 |
| nucleotide sequence of the major early region of bacteriophage phi 29. | the nucleotide sequence of the left end of bacteriophage phi 29 dna has been determined. together with data reported earlier (yoshikawa et al., 1981), this sequencing comprises the major early genetic region of this viral genome (5708 bp). computer analysis of the dna sequences revealed that there are up to fifteen open reading frames which could encode polypeptides containing more than thirty amino acids. the dna sequence also revealed a number of potential regulatory signals, promoters and rib ... | 1982 | 6809534 |
| initiation of phage phi 29 dna replication by mutants with deletions at the amino end of the terminal protein. | series of deletions at the amino end of protein p3, the phage phi 29 dna terminal protein (tp), have been constructed and characterized. measurements of the activity of the deletion mutants in the formation of the protein p3-damp initiation complex in vitro indicate the dispensability of the first 13 amino acids (aa) of the protein. the activity of protein p3 decreased considerably when 17 or more aa were deleted. the results on the in vitro phi 29 dna replication primed by the p3 deletion mutan ... | 1988 | 3133284 |
| symmetrical transcription in bacteriophage phi 29 dna. | transcription of some early genes occurring during phi 29 infection in the absence but not in the presence of chloramphenicol has been shown to depend upon the synthesis of the viral protein p4, the positive regulator of late transcription. in addition, the early promoter b1, responsible for early transcription on the late region of the phi 29 genome, has been accurately mapped by nuclease s1 protection experiments. the deduced promoter sequence shares homology with that of the other early phi 2 ... | 1988 | 3139079 |
| protein-primed replication of bacteriophage phi 29 dna. | the replication of phi 29 dna-protein p3 represents a simple model system to study the protein-priming mechanism of initiation of replication. the phi 29 dna polymerase involved both in the initiation and elongation steps of phi 29 dna-protein p3 replication, is a very processive enzyme and it is able to produce strand-displacement in the absence of other proteins. to correlate functional and structural domains in the phi 29 dna polymerase point mutants in the most carboxyl region of amino-acid ... | 1988 | 3207763 |
| adsorption of bacteriophage phi 29 to bacillus subtilis through the neck appendages of the viral particle. | phage phi 29 particles produced under restrictive conditions by mutants in gene 12 have normal amounts of all of the structural proteins except the appendage protein, p12*, which is missing. these particles are not infective and do not adsorb to bacillus subtilis cells. by in vitro complementation of 12- particles with extracts containing protein p12* or with purified protein p12*, the defective particles could bind the appendage protein and become infective and able to adsorb to bacteria. there ... | 1981 | 7241648 |
| dna replication of bacteriophage phi 29: characterization of the intermediates and location of the termini of replication. | | 1980 | 7395108 |
| a defined system for in vitro packaging of dna-gp3 of the bacillus subtilis bacteriophage phi 29. | the bacteriophage phi 29 dna-gene product 3 complex (dna-gp3) has been efficiently packaged into proheads in a completely defined in vitro system. the phi 29 dna packaging protein gp16, the product of gene 16, was overproduced in escherichia coli and purified to near homogeneity. the purified gp16 packaged 23% of the dna-gp3 added to purified proheads in the defined mixture, while gp16 in an extract of phage-infected cells packaged 26% of the dna-gp3. no host proteins were required in the define ... | 1986 | 3458193 |
| in vitro replication of bacteriophage phi 29. | | 1984 | 6395662 |
| in vitro assembly of infectious virions of double-stranded dna phage phi 29 from cloned gene products and synthetic nucleic acids. | up to 6 x 10(7) pfu of infectious virions of the double-stranded dna bacteriophage phi 29 per ml were assembled in vitro, with 11 proteins derived from cloned genes and nucleic acids synthesized separately. the genomic dna-gp3 protein conjugate was efficiently packaged into a purified recombinant procapsid with the aid of a small viral rna (prna) transcript, a dna-packaging atpase (gp16), and atp. the dna-filled capsids were subsequently converted into infectious virions after the addition of fo ... | 1995 | 7609071 |
| sequential interactions of structural proteins in phage phi 29 procapsid assembly. | the mechanism of viral capsid assembly is an intriguing problem because of its fundamental importance to research on synthetic viral particle vaccines, gene delivery systems, antiviral drugs, chimeric viruses displaying antigens or ligands, and the study of macromolecular interactions. the genes coding for the scaffolding (gp7), capsid (gp8), and portal vertex (gp10) proteins of the procapsid of bacteriophage phi 29 of bacillus subtilis were expressed in escherichia coli individually or in combi ... | 1995 | 7609072 |
| structural localization of the proteins of the head to tail connecting region of bacteriophage phi 29. | the head to tail connector of bacteriophage phi 29 has been studied to locate its two structural proteins (p10 and p11). treatment with trypsin led to proteolysis of p10 while p11 remained intact. computer filtration of electron micrographs of crystals of trypsinized necks showed a change in the external 12-fold area of the neck when compared with control necks. proteolized necks completely released p10 after treatment with low concentrations of an ionic detergent. the resulting structures, cont ... | 1983 | 6401885 |
| in vitro synthesis of late bacteriophage phi 29 rna. | a crude p-100 fraction prepared from bacillus subtilis 21 min after infection with wild-type phage phi 29 supported the in vitro synthesis of late phi 29 rna by added rna polymerase. synthesis of late rna was also detected when purified phi 29 dna was transcribed by rna polymerase in the presence of an s-150 fraction obtained by lysis of phi 29-infected cells in the presence of 1 m nacl. late phi 29 rna was not synthesized when either the p-100 or the s-150 fraction was prepared from cultures in ... | 1983 | 6406684 |
| protein-nucleic acid interactions in bacteriophage phi 29 dna replication. | phi 29 dna replication starts at both dna ends by a protein priming mechanism. the formation of the terminal protein-damp initiation complex is directed by the second nucleotide from the 3' end of the template. the transition from protein-primed initiation to normal dna elongation has been proposed to occur by a sliding-back mechanism that is necessary for maintaining the sequences at the phi 29 dna ends. structure-function studies have been carried out in the phi 29 dna polymerase. by site-dire ... | 1995 | 7669351 |
| characterization of an rna-binding domain in the bacteriophage phi 29 connector. | the connector of bacteriophage phi 29 is known to promote the viral prohead assembly, to bind dna, and to drive dna packaging into preformed viral shells in an rna-dependent process. in this report, the phi 29 connector protein, p10, is shown to bind rna in a sequence-independent fashion, and to possess an rna recognition motif comprised approximately the region between residues 21 and 94 of the p10 sequence. substitution mutants in specific amino acids of the rna-binding domain obtained by site ... | 1993 | 7690751 |
| a novel dna polymerase induced by bacillus subtilis phage phi 29. | a novel dna polymerase induced by bacillus subtilis bacteriophage phi 29 has been identified. this polymerase can be separated from host dna polymerase, by fractionation of extracts prepared from phage infected cells, using phosphocellulose chromatography. the isolated polymerase prefers poly(da)oligo(dt) as template. the dna polymerase isolated from the cells infected with a gene 2 temperature sensitive mutant (ts2) showed greater heat-lability than that induced by wild type phi 29. the ts2 dna ... | 1983 | 6424095 |
| a genetic approach to the identification of functional amino acids in protein p6 of bacillus subtilis phage phi 29. | protein p6 of the bacillus subtilis phage phi 29 is essential for in vivo viral dna replication. this protein activates the initiation of phi 29 dna replication in vitro by forming a multimeric nucleoprotein complex at the replication origins. the n-terminal region of protein p6 is involved in dna binding, as shown by in vitro studies with p6 proteins altered by deletions or missense mutations. we report on the development of an in vivo functional assay for protein p6. this assay is based on the ... | 1994 | 7808404 |
| dual translational start motif evolutionarily conserved in the holin gene of bacillus subtilis phage phi 29. | holins represent phage encoded lysis functions required for transit of the phage murein hydrolases to the periplasm. the lambda s, phage 21 s, and p22 13 holin genes contain a dual translational start motif, beginning with met1-lys2-x-met3. in all cases both start codons at the 5' end of the respective holin gene are utilized. the resulting polypeptides have opposing functions, with the longer product acting as an inhibitor of the shorter one. the 131-codon gene 14 of bacillus subtilis phage phi ... | 1995 | 7831803 |
| primary structure and functional analysis of the lysis genes of lactobacillus gasseri bacteriophage phi adh. | the lysis genes of the lactobacillus gasseri bacteriophage phi adh were isolated by complementation of a lambda sam mutation in escherichia coli. nucleotide sequencing of a 1,735-bp dna fragment revealed two adjacent coding regions of 342 bp (hol) and 951 bp (lys) in the same reading frame which appear to belong to a common transcriptional unit. proteins corresponding to the predicted gene products, holin (12.9 kda) and lysin (34.7 kda), were identified by in vitro and in vivo expression of the ... | 1995 | 7836307 |
| transcription regulation in bacillus subtilis phage phi 29: expression of the viral promoters throughout the infection cycle. | transcription of the genome of bacillus subtilis phage phi 29 is tightly controlled, taking place in two stages, early and late. we have analyzed the abundance of the transcripts produced from each viral promoter throughout the infection cycle. we compare the relative strength of each promoter, as well as get a better understanding of the regulatory events, finding a new promoter regulated by the viral protein p4. the two strong early promoters, a2b and a2c, responsible for the expression of gen ... | 1995 | 7871731 |
| purification in a functional form of the terminal protein of bacillus subtilis phage phi 29. | phage phi 29 terminal protein, p3, essentially pure, was isolated in a denatured form from viral particles, and anti-p3 antiserum was obtained. a radioimmunoassay to detect and quantitate protein p3 was developed. by using this assay, native protein p3 was highly purified from escherichia coli cells harboring a gene 3-containing recombinant plasmid. after three purification steps, the protein was more than 96% pure; its amino acid composition was very similar to that deduced from the nucleotide ... | 1984 | 6424120 |
| structure of bacteriophage phi 29 dna. | | 1984 | 6427030 |
| in vivo functional relationships among terminal proteins of bacillus subtilis phi 29-related phages. | gene 3 of the bacillus subtilis phage phi 29 encodes the terminal protein (tp), which acts as a primer in the initiation of viral dna replication. we have developed an in vivo functional assay for the phi 29 tp based on the ability of tp-producing b. subtilis non-suppressor (su-) cells to support dna replication of a phi 29 sus3 mutant phage. this trans-complementation assay has been used to study in vivo functional relationships between the tp of phi 29 and related phages. our results demonstra ... | 1994 | 7926823 |
| bacteriophage phi 29 proteins required for in vitro dna-gp3 packaging. | in vitro assembly of bacteriophage phi 29 in crude extracts involves efficient packaging of a dna-protein complex (dna- gp3 ) into a prohead with the aid of the gene 16 product ( gp16 ) and subsequent assembly of neck and tail proteins ( bjornsti et al., j. virol. 41:508-517, 1982; bjornsti et al., j. virol. 45:383-396, 1983; bjornsti et al., proc. natl. acad. sci. u.s.a. 78:5861-5865, 1981). to define the viral proteins required for the dna- gp3 encapsidation phase, we purified biologically act ... | 1984 | 6427474 |
| characterization and purification of a phage phi 29-encoded dna polymerase required for the initiation of replication. | the phage phi 29 protein p2, required for the formation of the protein p3-damp initiation complex, has been purified from escherichia coli cells harboring a gene 2-containing recombinant plasmid. the purified protein p2, of molecular weight 68,000, had a specific dna polymerase activity that elongated the p3-damp initiation complex when phi 29 dna-protein p3 was used as template. in addition, the purified protein p2 was active in catalyzing the initiation reaction when complemented with phi 29 m ... | 1984 | 6433348 |
| dna structure in the nucleoprotein complex that activates replication of phage phi 29. | initiation of phage phi 29 dna replication is activated by the viral protein p6 which forms a nucleoprotein complex at the replication origins, located at the linear genome ends. the complex consists of a dna right-handed superhelix wrapped around a multimeric protein core. we have determined the superhelical path of the dna in the complex, measuring the change in linking number induced by the protein, the surface-related helical repeat and the compaction of the dna. one superhelical turn has ap ... | 1994 | 8011933 |
| a highly sensitive system for the in vitro assembly of bacteriophage phi 29 of bacillus subtilis. | a sensitive system for the assay of bacteriophage phi 29 assembly in vitro was developed using 12 recombinant proteins and synthetic prna. this system detected in vitro assembled infectious phages up to 10(7) plaque forming units (pfu) per milliliter without any background. phi 29 dna-gp3 concentration dependence in phage assembly was found to be first order, while the dna-packaging protein gp16 dependence was higher order. the requirement for specific phi 29 prna for phi 29 dna packaging was co ... | 1994 | 8030206 |
| replication of bacteriophage phi 29 dna in vitro: the roles of terminal protein and dna polymerase. | phi 29 dna replication is initiated by the formation of a covalent complex between the viral-coded terminal protein and damp (tp-damp). this initiation reaction system has been reconstituted from two phage-encoded proteins, the terminal protein and dna polymerase. the phi 29 dna polymerase was purified from phage-infected cells by using poly(da) x p(dt)12-18 as an assay template. the purified polymerase has an apparent molecular mass of 68 kda in its native form and it appears to function as a m ... | 1984 | 6433349 |
| structural study of tetragonal-ordered aggregates of phage phi 29 necks. | a new class of two-dimensional tetragonal aggregates of phage phi 29 necks has been studied by electron microscopy and a combination of fourier filtering procedures and detailed rotational analysis. the results confirm the main features of the head-to-tail connecting region previously observed in hexagonal aggregates. there are several differences in the resulting pictures that can be attributed to the different way in which the aggregates are organized and stained. | 1984 | 6544883 |
| bacillus subtilis mutants defective in bacteriophage phi 29 head assembly. | virus assembly mutants of asporogenous bacillus subtilis defective in bacteriophage phi 29 head assembly were detected by the use of antibodies that reacted strongly with the free dodecameric phi 29 portal vertex composed of gene product 10 (gp10) but weakly with the portal vertex assembled into proheads or phage. phage adsorption and the synthesis of phage proteins, dna-gene product 3, and prohead rna were normal in these mutants, but prohead and phage production was greatly reduced. the assemb ... | 1993 | 8096839 |
| protein p3 is linked to the dna of phage phi 29 through a phosphoester bond between serine and 5'-damp. | to investigate the role of protein p3 in bacteriophage phi 29 initiation of replication, we have studied the nature of the covalent linkage between protein p3 and phi 29 dna. the protein-dna compound was digested with micrococcal nuclease and pronase resulting in a nucleotidyl-peptide that was further digested by alkaline phosphatase and snake venom phosphodiesterase yielding 5'-damp. the dna-protein linkage is sensitive to alkali. treatment of the nucleotidyl-peptide with 0.1 m naoh at 37 degre ... | 1980 | 6779279 |
| bacteriophage phi 29 infection of bacillus subtilis minicells. | bacteriophage phi 29 can infect b. subtilis minicells and synthesize all the phage-coded proteins detected in ultraviolet irradiated-infected b. subtilis cells. synthesis of phage unit-length dna has been obtained after infection of minicells with phi 29. the dna can be encapsulated in particles with a sedimentation rate similar to that of phage phi 29 produced in b. subtilis cells. the particles produced in minicells can be adsorbed to b. subtilis cells, but infectivity has not been demonstrate ... | 1980 | 6780760 |
| purification, properties and assembly of the neck-appendage protein of the bacillus subtilis phage phi 29. | the purification of the neck appendage protein of phi 29, p12*, which is involved in the adsorption of the phage to bacillus subtilis, is described. the purified native protein is in a dimeric form and can be assembled, in vitro, onto purified 12- particles that lack the neck appendages, suggesting that the incorporation of p12* to the rest of the phage structure is a self-assembly process. the assembly of protein p12* in vitro follows cooperative kinetics and it occurs with an efficiency of abo ... | 1981 | 6793359 |
| initiation factor-independent translation of mrnas from gram-positive bacteria. | initiation factor-independent translation of mrna derived from bacillus phage phi29 dna occurs with translation systems derived from bacillus subtilis or escherichia coli. this is in sharp contrast to the strict dependence on ribosome salt wash fraction of e. coli ribosomes for the translation of t7 and other mrnas derived from gram-negative organisms. | 1981 | 6795625 |
| in vitro assembly of the bacillus subtilis bacteriophage phi 29. | in vitro assembly of the bacillus subtilis bacteriophage phi 29 that approaches the efficiency of assembly in vivo has been demonstrated. proheads, dna, and gene 16 product (gp16) were essential for dna encapsidation, and the average yield in extracts was 180 phage per prohead donor cell. the in vitro maturation was very similar to in vivo assembly in terms of yield, intermediates, and abortive structures. more that 30% of the proheads in the extract were converted to phage, and about 20% of dna ... | 1981 | 6795639 |
| rna-mediated specificity of dna packaging into hybrid lambda/phi 29 proheads. | a small rna (prna, 174 nt) is known to be essential for dna packaging in bacteriophage phi 29. however, in an in vitro dna packaging system based on hybrid lambda/phi 29 proheads (made up of head proteins from phage lambda and connectors from phage phi 29), the specificity of dna packaging is lost, and different rna molecules fulfil the requirements for dna packaging, albeit with less efficiency than phi 29 prna. competition assays with rnas from different sources have shown that phi 29 connecto ... | 1993 | 8223455 |
| structure of the head-tail connector of bacteriophage phi 29. | | 1982 | 6804634 |
| in vitro complex formation between bacteriophage phi 29 terminal protein and deoxynucleotide. | | 1982 | 6807309 |
| nucleotide sequence of the early genes 3 and 4 of bacteriophage phi 29. | the nucleotide sequence of an early region of the phi 29 genome has been determined. the sequenced region includes genes 3 and 4, which code for the protein covalently linked to the 5' ends of phi 29 dna and the protein involved in the control of late transcription, respectively. the position and nature of the mutations of mutants sus3(91) and sus4(56) has also been determined. | 1982 | 6292852 |