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PMID: 6294482 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

On the role of the single-stranded DNA binding protein of bacteriophage T4 in DNA metabolism. I. Isolation and genetic characterization of new mutations in gene 32 of bacteriophage T4.

Molecular & general genetics : MGG ·Vol. 188 ·No. 1 ·1982-00-00 ·Pages 77-90

Doherty DH, Gauss P, Gold L

Abstract

The product of gene 32 of bacteriophage T4 is a single-stranded DNA binding protein involved in T4 DNA replication, recombination and repair. Functionally differentiated regions of the gene 32 protein have been described by protein chemistry. As a preliminary step in a genetic dissection of these functional domains, we have isolated a large number of missense mutants of gene 32. Mutant isolation was facilitated by directed mutagenesis and a mutant bacterial host which is unusually restrictive for missense mutations in gene 32. We have isolated over 100 mutants and identified 22 mutational sites. A physical map of these sites has been constructed and has shown that mutations are clustered within gene 32. The possible functional significance of this clustering is considered.

MeSH Terms
Base Sequence DNA Helicases/metabolism DNA, Recombinant DNA, Single-Stranded/metabolism DNA, Viral/metabolism DNA-Binding Proteins Escherichia coli Genes, Viral Mutation T-Phages/genetics
Chemicals
DNA, Recombinant DNA, Single-Stranded DNA, Viral DNA-Binding Proteins DNA Helicases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Doherty D H
Gauss P
Gold L
References (50)
50 references, click to expand
  1. Molecular recombination in T4 bacteriophage deoxyribonucleic acid. II. Single-strand breaks and exposure of uncomplemented areas as a prerequisite for recombination.
    J Virol. 1967 Dec;1(6):1193-202 PMID: 4912244
  2. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form.
    Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159-66 PMID: 4894690
  3. rII cistrons of bacteriophage T4. DNA sequence around the intercistronic divide and positions of genetic landmarks.
    J Mol Biol. 1981 Jul 5;149(3):337-76 PMID: 6273585
  4. UGA: a third nonsense triplet in the genetic code.
    Nature. 1967 Feb 4;213(5075):449-50 PMID: 6032223
  5. Construction and properties of a recombinant plasmid containing gene 32 of bacteriophage T4D.
    J Mol Biol. 1981 May 25;148(3):199-218 PMID: 6271969
  6. Two types of replication proteins increase the rate at which T4 DNA polymerase traverses the helical regions in a single-stranded DNA template.
    J Biol Chem. 1981 Apr 25;256(8):4087-94 PMID: 6971292
  7. Characterization of a bacteriophage T4 mutant lacking DNA-dependent ATPase.
    J Virol. 1975 Jan;15(1):50-4 PMID: 123009
  8. Multiple interactions of a DNA-binding protein in vivo. III. Phage T4 gene-32 mutations differentially affect insertion-type recombination and membrane properties.
    Genetics. 1977 May;86(1):5-23 PMID: 885343
  9. Genetic identification of cloned fragments of bacteriophage T4 DNA and complementation by some clones containing early T4 genes.
    Mol Gen Genet. 1977 Sep 9;154(3):319-26 PMID: 927440
  10. DNA replication by bacteriophage T4 proteins. The T4 43, 32, 44--62, And 45 proteins are required for strand displacement synthesis at nicks in duplex DNA.
    J Biol Chem. 1979 Jul 10;254(13):6032-7 PMID: 376525
  11. Physiochemical properties of DNA binding proteins: gene 32 protein of T4 and Escherichia coli unwinding protein.
    Biochemistry. 1975 Dec 16;14(25):5485-91 PMID: 1103969
  12. T4 bacteriophage gene 32: a structural protein in the replication and recombination of DNA.
    Nature. 1970 Sep 26;227(5265):1313-8 PMID: 5455134
  13. Mutants of bacteriophage T4 which allow amber mutants of gene 32 to grow in ochre-suppressing hosts.
    Virology. 1973 May;53(1):47-59 PMID: 4574876
  14. Requirement of a functional gene 32 product of bacteriophage T4 in UV, repair.
    J Virol. 1973 Oct;12(4):758-65 PMID: 4591048
  15. Translational, autogenous regulation of gene 32 expression during bacteriophage T4 infection.
    J Biol Chem. 1976 Nov 25;251(22):7263-70 PMID: 791948
  16. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9.
    Gene. 1977;2(2):75-93 PMID: 344136
  17. Coupling of late transcription to viral replication in bacteriophage T4 development.
    J Mol Biol. 1970 Nov 28;54(1):85-102 PMID: 4923869
  18. Physical chemical studies of the structure and function of DNA binding (helix-destabilizing) proteins.
    CRC Crit Rev Biochem. 1980 Jan;7(3):247-89 PMID: 6243083
  19. The UV sensitivity of some early-function temperature-sensitive mutants of phage T4.
    Virology. 1970 Feb;40(2):272-87 PMID: 4909413
  20. Variations in genetic recombination due to amber mutations in T4D bacteriophage.
    J Virol. 1969 Feb;3(2):171-5 PMID: 4886654
  21. Induction of mutations in specific genes of bacteriophage T4 using cloned restriction fragments and marker rescue.
    Mol Gen Genet. 1980 Feb;177(3):447-52 PMID: 6990202
  22. Studies with DNA-cellulose chromatography. I. DNA-binding proteins from Escherichia coli.
    Cold Spring Harb Symp Quant Biol. 1968;33:289-305 PMID: 4891970
  23. Interactions of bacteriophage T4-coded gene 32 protein with nucleic acids. I. Characterization of the binding interactions.
    J Mol Biol. 1981 Jan 5;145(1):75-104 PMID: 7265204
  24. DNA helicases.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:63-7 PMID: 225120
  25. Genetic evidence for an additional function of phage T4 gene 32 protein: interaction with ligase.
    Proc Natl Acad Sci U S A. 1975 Apr;72(4):1226-30 PMID: 1055398
  26. The identification of complex genotypes in bacteriophage T4. I. Methods.
    Genetics. 1970 Nov;66(3):417-28 PMID: 4933468
  27. A strong suppressor specific for UGA.
    Nature. 1967 Apr 29;214(5087):452-3 PMID: 5340340
  28. DNA "melting" proteins. II. Effects of bacteriophage T4 gene 32-protein binding on the conformation and stability of nucleic acid structures.
    J Biol Chem. 1976 Nov 25;251(22):7215-28 PMID: 791945
  29. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid.
    J Bacteriol. 1978 Jun;134(3):1141-56 PMID: 149110
  30. Regulation of gene 32 expression during bacteriophage T4 infection of Escherichia coli.
    J Biol Chem. 1976 Nov 25;251(22):7251-62 PMID: 791947
  31. Gene 32 protein of bacteriophage T4 moderates the activities of the T4 gene 46/47-controlled nuclease and of the Escherichia coli RecBC nuclease in vivo.
    J Virol. 1976 Mar;17(3):756-61 PMID: 1255856
  32. Structure of gene 5 protein-oligodeoxynucleotide complexes as determined by 1H, 19F, and 31P nuclear magnetic resonance.
    Biochemistry. 1976 Dec 14;15(25):5419-30 PMID: 999816
  33. Probing DNA replication mechanisms with the T4 bacteriophage in vitro system.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:469-87 PMID: 225114
  34. Regulation of the synthesis of bacteriophage T4 gene 32 protein.
    J Mol Biol. 1974 Sep 5;88(1):89-104 PMID: 4444040
  35. Mutations affecting genetic recombination in bacteriophage T4D. I. Pathway analysis.
    Virology. 1977 Jul 1;80(1):67-82 PMID: 878316
  36. Sequences of the ssb gene and protein.
    Proc Natl Acad Sci U S A. 1981 Jul;78(7):4274-8 PMID: 6270666
  37. Molecular mechanisms of genetic recombination in bacteriophage: joint molecules and their conversion to recombinant molecules.
    J Cell Physiol. 1967 Oct;70(2):Suppl:201-13 PMID: 5582806
  38. Mutations altering genetic recombination and repair of DNA in bacteriophage T4.
    Virology. 1975 Feb;63(2):539-67 PMID: 163533
  39. Structure at 2.3 A resolution of the gene 5 product of bacteriophage fd: a DNA unwinding protein.
    J Mol Biol. 1979 Nov 5;134(3):379-400 PMID: 231669
  40. Primary structure of the bacteriophage T4 DNA helix-destabilizing protein.
    J Biol Chem. 1981 Feb 25;256(4):1754-62 PMID: 6257686
  41. In vivo interactions of genes and proteins in DNA replication and recombination of phage T4.
    Cold Spring Harb Symp Quant Biol. 1979;43 Pt 1:501-15 PMID: 289460
  42. Ribosomal mutations affecting efficiency of amber suppression.
    J Mol Biol. 1970 Feb 14;47(3):517-30 PMID: 4907270
  43. Autogenous translational repression of bacteriophage T4 gene 32 expression in vitro.
    J Mol Biol. 1978 Nov 25;126(1):73-90 PMID: 739544
  44. Interactions of bacteriophage T4-coded gene 32 protein with nucleic acids. III. Binding properties of two specific proteolytic digestion products of the protein (G32P*I and G32P*III).
    J Mol Biol. 1981 Jan 5;145(1):123-38 PMID: 6455528
  45. Interactions of bacteriophage T4-coded gene 32 protein with nucleic acids. II. Specificity of binding to DNA and RNA.
    J Mol Biol. 1981 Jan 5;145(1):105-21 PMID: 7265197
  46. Nucleotide sequence from the genetic left end of bacteriophage T7 DNA to the beginning of gene 4.
    J Mol Biol. 1981 Jun 5;148(4):303-30 PMID: 7310871
  47. Stimulation of T4 bacteriophage DNA polymerase by the protein product of T4 gene 32.
    J Mol Biol. 1971 Nov 28;62(1):39-52 PMID: 4945535
  48. A mutation that confers temperature sensitivity on the translation of rIIB in bacteriophage T4.
    J Mol Biol. 1976 May 25;103(3):627-46 PMID: 940158
  49. Branched DNA molecules: intermediates in T4 recombination.
    J Mol Biol. 1971 Aug 28;60(1):131-49 PMID: 4937189
  50. Mutagenic treatment of double- and single-stranded DNA phages T4 ans S13 with hydroxylamine.
    Virology. 1968 Jun;35(2):330-3 PMID: 5663335
Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00332998
Published
1982-00-00
Pages
77-90
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
Grants
NIGMS NIH HHS · GM06459 · United States
NIGMS NIH HHS · GM19963 · United States
External Links
PubMed source DOI: 10.1007/BF00332998
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