Summary
We have previously identified a unique site, pac, from which packaging of precursor concatameric viral DNA into proheads starts during the maturation process of bacteriophage CP-T1. The direction of this packaging was determined from restriction enzyme cleavage patterns of CP-T1 DNA. A restriction enzyme generated fragment containing pac was cloned and the surrounding DNA region sequenced. Analysis of the nucleotide sequence revealed numerous repeat regions related to the consensus sequence PuagttGAT.AAT.aa.t. Within the sequenced region an open reading frame encoding a 12260 Mr protein was also identified. This protein appears to share homology with the binding domains of known DNA binding proteins and may represent a putative Pac terminase possessing the specific endonuclease activity required for cleavage at the pac site. Minicell analysis of deletion derivatives of the pac-containing clone revealed a protein of approximately 12900 Mr encoded within this same region, confirming that this Pac protein is phage encoded.
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References
Achtman M, Schwuchow S, Helmuth R, Morelli G, Manning PA (1978) Cell-cell interactions in conjugating Escherichia coli: Con- mutants and stabilization of mating aggregates. Mol Gen Genet 164:171–183
Achtman M, Manning PA, Edelbluth C, Herrlich P (1979) Export without proteolytic processing of inner and outer membrane proteins encoded by F sex factor tra cistrons in Escherichia coli minicells. Proc Natl Acad Sci USA 76:4837–4841
Argos P (1985) Evidence for a repeating domain in Type I restriction enzymes. EMBO J 4:1351–1355
Bächi B, Arber W (1977) Physical mapping of BglII, BamHI, EcoRI, HindIII and PstI restriction fragments of bacteriophage P1 DNA. Mol Gen Genet 153:311–324
Backhaus H (1985) The packaging initiation of Salmonella bacteriophage P22: determination of cut sites within the DNA sequence coding for gene 3. J Virol 55:458–465
Becker A, Gold M (1978) Enzymatic breakage of the cohesive end site of phage λ DNA: terminase (ter) reaction. Proc Natl Acad Sci USA 75:4199–4203
Behnisch W, Schmieger H (1985) In vitro packaging of plasmid DNA oligomers by Salmonella phage P22: independence of the pac site and evidence for the termination cut in vitro. Virology 144:310–317
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneckes HL, Boyer HW (1977) Construction and characterization of new cloning vehicles. II. A multi-purpose cloning system. Gene 2:95–113
Casjens S, Huang WM (1982) Initiation of sequential packaging of bacteriophage P22 DNA. J Mol Biol 157:287–298
Chou PY, Fasman GD (1978) Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol 47:45–148
Deichelbohrer I, Messer W, Truatner TA (1982) Genome of Bacillus subtilis bacteriophage SPP1 structure and nucleotide sequence of pac the origin of DNA packaging. J Virol 42:83–90
Feiss M, Siegele DA,(1979) Packaging of the bacteriophage lambda chromosome: dependence of cos cleavage on chromosome length. Virology 92:190–200
Feiss M, Widner W (1982) Bacteriophage λ DNA packaging: scanning for the terminal cohesive end site during packaging. Proc Natl Acad Sci USA 79:7498–3502
Gicquel-Sanzey B, Cossart P (1982) Homologies between different procaryotic DNA-binding regulatory proteins and between their sites of action. EMBO J 1:591–595
Groenen MAM, Van de Putte P (1985). Mapping of a site for packaging of bacteriophage Mu DNA. Virology 144:520–522
Guidolin A, Morelli G, Kamke M, Manning PA (1984) Bacteriophage CP-T1 of Vibrio cholerae: characterization of phage DNA and restriction analysis. J Virol 51:163–169
Jackson EN, Miller HJ, Adams JC (1978) EcoRI restriction endonuclease site map of bacteriophage P22 DNA. J Mol Biol 118:347–363
Jackson EN, Laski F, Andres C (1982) Bacteriophage P22 mutants that alter the specificity of DNA packaging. J Mol Biol 154:551–563
Kennedy N, Beutin L, Achtman M, Skurray R, Rahmsdorf U, Herrlich P (1977) Conjugation proteins of the F sex factor. Nature 270:580–585
Kufer B, Backhaus H, Schmieger H (1982) The packaging initiation site of phage P22, analysis of packaging events by transduction. Mol Gen Genet 187:510–515
Laski F, Jackson E (1982) Maturation cleavage of bacteriophage P22 DNA in the absence of DNA packaging. J Mol Biol 154:565–580
Lugtenberg B, Meijers J, Peters R, van der Hoek P, van Alphen L (1975) Electrophoretic resolution of the ‘major outer membrane protein’ of Escherichia coli into four bands. FEBS Lett 58:254–258
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning — A laboratory manual. Cold Spring Harbor Laboratory, NY
McClelland M (1983) The effect of site specific methylation on restriction endonuclease cleavage (update). Nucleic Acids Res 11:r169-r173
Messing J, Vieira J (1982) A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene 19:269–276
Morelli G, Fisseau C, Behrens B, Trautner TA, Luh J, Ratcliff SW, Allison DP, Ganesan AT (1979) The genome of B. subtilis phage SPP1: the topology of DNA molecules. Mol Gen Genet 168:153–164
Mousset S, Thomas R (1969) ter a function which generates the ends of the mature λ chromosome. Nature 221:242–244
Pabo CO, Sauer RT (1984) Protein — DNA recognition. Annu Rev Biochem 53:293–331
Ratcliff SW, Luh J, Ganesan AT, Behrens B, Thompson R, Montenegro M, Morelli G, Trautner TA (1979) The genome of B. subtilis phage SPP1: the arrangement of restriction endonuclease generated fragments. Mol Gen Genet 168:165–172
Sangen F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sanger F, Coulson AR, Barrell BG, Smith AJH, Roe BA (1980) Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing. J Mol Biol 143:161–178
Schmieger H (1984) pac sites are indispensible for in vitro packaging of DNA by phage P22. Mol Gen Genet 195:252–255
Sternberg N, Wieisberg R (1977) Packaging of coli-phage lambda DNA. I. The role of the cohesive end site and the gene A protein. J Mol Biol 117:717–731
Streisinger G, Emrick J, Stahl MM (1967) Chromosome structure in phage T4. III. Terminal redundancy and length determination. Proc Natl Acad Sci USA 57:292–295
Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171
Tye BK, Huberman JA, Botstein D (1974) Non-randon circular permutation of phage P22 DNA. J Mol Biol 85:501–532
Wang J, Kaiser AD (1973) Evidence that cohesive ends of mature DNA are generated by the gene A product. Nature 241:16–17
Yamagishi M, Fujisawa H, Minagawa T (1985) Isolation and characterization of bacteriophage T3/T7 hybrids and their use in studies on molecular basis of DNA packaging specificity. Virology 144:502–515
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Guidolin, A., Manning, P.A. Molecular analysis of the packaging signal in bacteriophage CP-T1 of Vibrio cholerae . Mol Gen Genet 212, 514–521 (1988). https://doi.org/10.1007/BF00330858
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DOI: https://doi.org/10.1007/BF00330858