a transcriptional map of the bacteriophage sp01 genome: ii. the major early transcription units.the genome of the bacillus subtilis phage sp01 contains a 12.6-kilobase-pair terminal redundancy. the transcription units in this region, which are utilized in vitro by b. subtilis rna polymerase holoenzme, have been mapped. in vitro transcripts were separated and characterized by gel electrophoresis. since there is such a large number of promoters in this region, it was necessary to employ conditions of transcription under which only subsets of all the transcripts would be made. selective synth ...198118635060
limited role of parental dna in replication during infection by bacillus subtilis phage spo1.when bacteria, growing in light medium, are infected with heavy phage, progeny dna molecules band in the hybrid position only if they were synthesized using parental dna as template. thus, the rate of incorporation of radioactive precursors into hybrid dna measures the use of parental template. this rate decreases with time during spo1 infection, arguing against certain models for the control of spot replication.198118635046
inhibitory action of erythromycin on bacteriophage spo1 multiplication in sporulating cells of bacillus subtilis 168.erythromycin (2--4 microgram/ml) was found to inhibit specifically multiplication of spo1 in sporulating cells of an erythromycin-resistant, conditional asporogenous mutant of bacillus subtilis 168 thy- trp-, ery1040. in contrast, streptomycin (150--200 microgram/ml) which inhibits protein synthesis to a similar extent as erythromycin did not inhibit spo1 multiplication severely, suggesting that the inhibition of spo1 multiplication by erythromycin is not caused by an overall inhibition of prote ...19806777627
[bacillus subtilis mutant with altered ability for heterologous transformation].mutant strain of bacillus subtilis, which produced in certain conditions significantly reduced quantity of trnasformants during transformation by homologous dna, as compared with transformation by heterologous dna from bac. aterrimus, is isolated. the ability to transfection by phage spo1 dna and the efficiency of infection of the mutant by this phage are also decreased. the causes of such alterated properties are discussed.1978414961
definitive identification of mammalian 5-hydroxymethyluracil dna n-glycosylase activity as smug1.purification from calf thymus of a dna n-glycosylase activity (hmudg) that released 5-hydroxymethyluracil (5hmura) from the dna of bacillus subtilis phage spo1 was undertaken. analysis of the most purified fraction by sds-polyacrylamide gel electrophoresis revealed a multiplicity of protein species making it impossible to identify hmudg by inspection. therefore, we renatured the enzyme after sds-polyacrylamide gel electrophoresis and assayed slices of the gel for dna n-glycosylase activity direc ...200111526119
genes that protect against the host-killing activity of the e3 protein of bacillus subtilis bacteriophage spo1.a cloned rpob gene, specifying an apparently mutant rna polymerase beta subunit, protected escherichia coli against the cytocidal effects of the e3 protein of bacteriophage spo1, suggesting that rna polymerase is the primary cellular target of the e3 protein. two segments of the wild-type e. coli genome, one of which specifies a suppressor of dnak mutations, and thus, possibly, a molecular chaperone, also provided protection when overexpressed, but wild-type rpob did not.19957751311
bacteriophage spo1 gene 27: location and nucleotide sequence.bacteriophage spo1 gene 27, whose product is required for late gene transcription and dna replication, has been cloned in escherichia coli, and its complete nucleotide sequence has been determined. we infer that the product of gene 27 is a highly basic 17,518-dalton protein of 155 amino acids. the gene for this regulatory protein is transcribed from two promoters: an early promoter situated before the adjacent upstream gene 28 and a middle promoter located between genes 28 and 27.19836413701
purification and dna-binding properties of rna polymerase from bacillus subtilis.four rna-polymerizing activities having different subunit composition can be purified from uninfected and from spo1-infected bacillus subtilis. lysozyme and sodium deoxycholate are used for lysing the cells. polymin p is used for precipitating nucleic acids and deae-cellulose chromatography allows separation of enzymatic activity from the residual polymin p. after these common steps, one can purify core + sigma + delta by chromatography on single-stranded dna-agarose followed by gel filtration w ...19806772439
structure of a bacillus subtilis bacteriophage spo1 gene encoding rna polymerase sigma factor.gene 28 of bacillus subtilis bacteriophage spo1 codes for a regulatory protein, a sigma factor known as sigma gp28, that binds to the bacterial core rna polymerase to direct the recognition of phage middle gene promoters. middle promoters exhibit distinctive and conserved nucleotide sequences in two regions centered about 10 and 35 base pairs upstream from the start point of mrna synthesis. here we report the cloning of gene 28 and its complete nucleotide sequence. we infer that sigma gp28 is a ...19836402778
effects of the positively regulating product of gene 28 of the b. subtilis phage spo1 on in vitro transcription.some of the properties of the rna polymerase purified from spo1-infected bacillus subtilis have been compared with the properties of rna polymerase from uninfected cells (core + sigma). the two enzymes synthetize rna from nonoverlapping regions on spo1 dna, and they lead to the retention of different restriction fragments of spo1 dna on cellulose-nitrate filters. the action of the positively regulating product of gene 28 of spo1 (gp 28) has been analyzed. the isolated gp 28 has been shown to be ...19816174844
new phage-spo1-induced polypeptides associated with bacillus subtilis rna polymerase.rna polymerase was precipitated from extracts of bacillus subtilis infected with phage sp01 by antiserum prepared against core rna polymerase. as shown by sodium dodecyl sulfate gel electrophoresis, the precipitates contained at least five new polypeptides not present in uninfected bacteria, in addition to the known subunits of rna polymerase. the molecular weights of these polypeptides are (1) 85,000; (ii) 40,000; (iii) 28,000; (iv) 25,000; and (v) 23,000. four of the polypeptides (i, iii, iv, ...19744212197
activity of two strong promoters cloned into bacillus subtilis.two dna fragments, one encoding the escherichia coli trc promoter and the other encoding a sequence from the early region of bacillus subtilis phage spo1, were cloned into the b. subtilis promoter-probe vector ppl603. both fragments effected strong in vivo promoter activity in vegetative b. subtilis cells.19863086500
transcription specificity of an rna polymerase fraction from bacteriophage sp01-infected b. subtilis. 19734201173
sequence of the bacteriophage sp01 gene coding for transcription factor 1, a viral homologue of the bacterial type ii dna-binding proteins.the bacillus subtilis phage sp01, whose dna contains 5-hydroxymethyluracil (hmura) in place of thymine, codes for an abundant, small, basic protein called tf1. tf1 binds preferentially to hydroxymethyluracil-containing dna and thereby selectively inhibits transcription of such dna in vitro. the gene for tf1 has been sequenced. we find that this viral protein is a homologue of the ubiquitous bacterial type ii dna-binding proteins. the three-dimensional structure of one of these bacterial proteins ...19846438630
the dna polymerase-encoding gene of bacillus subtilis bacteriophage spo1.the bacteriophage spo1 dna polymerase-encoding gene, which contains a self-splicing intron, has been sequenced and its amino acid (aa) sequence has been deduced. the aa sequence of spo1 dna polymerase shows a high degree of similarity with that of dna polymerase i from escherichia coli (po1i). alignment with the sequences of po1i, and the phi 29 and spo1 dna polymerases indicate that the aa residues that have been implicated in 3'----5' exonuclease activities are conserved.19921324872
the genetics of bacteriophage spo1. 19724628367
interaction of bacillus subtilis rna polymerase core with two specificity-determining subunits. competition between sigma and the spo1 gene 28 protein.gene activity in the development of phage spo1 is transcriptionally regulated. early viral genes are transcribed by the major vegetative cell rna polymerase (e. sigma). middle viral genes are transcribed by rna polymerase core (e) bearing the spo1 gene 28 protein (gp28) instead of sigma. this paper deals with the competitive interactions of sigma and gp28 with e which must, at least in part, be involved in the ability of viral middle gene expression to succeed early gene expression. an in vitro ...19826281274
stoichiometry of dna binding by the bacteriophage sp01-encoded type ii dna-binding protein tf1.the stoichiometry of dna binding by the bacteriophage sp01-encoded type ii dna-binding protein tf1 has been determined. 3h-labeled tf1 was allowed to bind to a 32p-labeled dna fragment containing a tf1 binding site. multiple tf1-dna complexes were resolved from each other and from unbound dna by native gel electrophoresis. dna-protein complexes were cut from polyacrylamide gels, and the amounts of 3h and 32p contained in each slice were measured. a ratio of 1.12 +/- 0.06 tf1 dimer/dna molecule w ...19902113049
bacteriophage spo1 dna- and rna-directed protein synthesis in vitro: comparison with in vivo control.a cell free protein synthesizing system, derived from e. coli, is shown to be a quantitiative assay system for messenger rna extracted from b. subtilis infected with bacteriophage spo1. dna-directed protein synthesis in this system is shown to be limited mostly to those proteins whose messages are contained in early rna. a phage induced enzyme, dcmp deaminase, is shown to be dependent on appearance of a class mrna made in vivo in response to new initiations of transcription dependent on prior sy ...1975810656
tf1, the bacteriophage spo1-encoded type ii dna-binding protein, is essential for viral multiplication.the lytic bacillus subtilis bacteriophage spo1 encodes an abundant, 99-amino-acid type ii dna-binding protein, transcription factor 1 (tf1). tf1 is special in this family of procaryotic chromatin-forming proteins in its preference for hydroxymethyluracil-containing dna, such as spo1 dna, and in binding with high affinity to specific sites in the spo1 chromosome. we constructed recessive null alleles of the tf1 gene and introduced them into spo1 chromosomes. segregation analysis with partially di ...19882841496
rna synthesis during bacteriophage spo1 development: six classes of spo1 rna. 19714996233
a self-splicing group i intron in the dna polymerase gene of bacillus subtilis bacteriophage spo1.we report a self-splicing intron in bacteriophage spo1, whose host is the gram-positive bacillus subtilis. the intron contains all the conserved features of primary sequence and secondary structure previously described for the group ia introns of eukaryotic organelles and the gram-negative bacteriophage t4. the spo1 intron contains an open reading frame of 522 nucleotides. as in the t4 introns, this open reading frame begins in a region that is looped out of the secondary structure, but ends in ...19902119891
fluorescence studies of a single tyrosine in a type ii dna binding protein.we studied the fluorescence properties of a single tyrosine (tyr94) located in the c-terminal tail of transcription factor 1 (tf1), a type ii procaryotic dna binding protein encoded by the bacillus subtilis phage spo1. the time-resolved fluorescence intensity of tyr94 in free tf1 dimers decays as a single exponential, and this is consistent with a twofold symmetrical structure. the fluorescence is readily quenched by acrylamide, but it is less accessible to anionic quenchers (iodide and citrate) ...19892706265
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