Skip to main content
Log in

Molecular analysis of the packaging signal in bacteriophage CP-T1 of Vibrio cholerae

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

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

    Google Scholar 

  • 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

    Google Scholar 

  • Argos P (1985) Evidence for a repeating domain in Type I restriction enzymes. EMBO J 4:1351–1355

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • Casjens S, Huang WM (1982) Initiation of sequential packaging of bacteriophage P22 DNA. J Mol Biol 157:287–298

    Google Scholar 

  • Chou PY, Fasman GD (1978) Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol 47:45–148

    Google Scholar 

  • 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

    Google Scholar 

  • Feiss M, Siegele DA,(1979) Packaging of the bacteriophage lambda chromosome: dependence of cos cleavage on chromosome length. Virology 92:190–200

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • Groenen MAM, Van de Putte P (1985). Mapping of a site for packaging of bacteriophage Mu DNA. Virology 144:520–522

    Google Scholar 

  • 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

    Google Scholar 

  • Jackson EN, Miller HJ, Adams JC (1978) EcoRI restriction endonuclease site map of bacteriophage P22 DNA. J Mol Biol 118:347–363

    Google Scholar 

  • Jackson EN, Laski F, Andres C (1982) Bacteriophage P22 mutants that alter the specificity of DNA packaging. J Mol Biol 154:551–563

    Google Scholar 

  • Kennedy N, Beutin L, Achtman M, Skurray R, Rahmsdorf U, Herrlich P (1977) Conjugation proteins of the F sex factor. Nature 270:580–585

    Google Scholar 

  • 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

    Google Scholar 

  • Laski F, Jackson E (1982) Maturation cleavage of bacteriophage P22 DNA in the absence of DNA packaging. J Mol Biol 154:565–580

    Google Scholar 

  • 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

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning — A laboratory manual. Cold Spring Harbor Laboratory, NY

    Google Scholar 

  • McClelland M (1983) The effect of site specific methylation on restriction endonuclease cleavage (update). Nucleic Acids Res 11:r169-r173

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • Mousset S, Thomas R (1969) ter a function which generates the ends of the mature λ chromosome. Nature 221:242–244

    Google Scholar 

  • Pabo CO, Sauer RT (1984) Protein — DNA recognition. Annu Rev Biochem 53:293–331

    Google Scholar 

  • 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

    Google Scholar 

  • Sangen F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467

    Google Scholar 

  • 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

    Google Scholar 

  • Schmieger H (1984) pac sites are indispensible for in vitro packaging of DNA by phage P22. Mol Gen Genet 195:252–255

    Google Scholar 

  • 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

    Google Scholar 

  • 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

    Google Scholar 

  • Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171

    Google Scholar 

  • Tye BK, Huberman JA, Botstein D (1974) Non-randon circular permutation of phage P22 DNA. J Mol Biol 85:501–532

    Google Scholar 

  • Wang J, Kaiser AD (1973) Evidence that cohesive ends of mature DNA are generated by the gene A product. Nature 241:16–17

    Google Scholar 

  • 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

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by W. Goebel

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00330858

Key words

Navigation