Bacteriophage T7 gene 2.5 protein: an essential protein for DNA replication.

Kim, Young Tae; Richardson, Charles C.

doi:10.1073/pnas.90.21.10173

Cited by 63 publications

(62 citation statements)

References 49 publications

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“…However, at least in the case of the T7/E. coli system, the phage ssDNA-binding protein is essential for infection (40), demonstrating that the host ssDNA-binding protein cannot substitute for that encoded by the phage. Most likely the ability of the phage ssDNA-binding protein to participate in phage-specific proteinprotein interactions confers its indispensable function.…”

Section: Discussionmentioning

confidence: 99%

Acidic C-terminal tail of the ssDNA-binding protein of bacteriophage T7 and ssDNA compete for the same binding surface

Marintcheva

Marintchev

Wagner

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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Section: Discussionmentioning

confidence: 99%

Acidic C-terminal tail of the ssDNA-binding protein of bacteriophage T7 and ssDNA compete for the same binding surface

Marintcheva

Marintchev

Wagner

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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“…A rate-limiting step in the phage life cycle can be the accumulation of a sufficient pool of nucleoside triphosphates to be incorporated into newly synthesized DNA. Phage T7 has partially solved this problem by using the dNMPs derived from host DNA, thus bypassing the complicated step of reduction of ribonucleotides (2,3). If any step were to be rate-limiting in T7-infected cells, then one might choose the conversion of nucleoside monophosphates to nucleoside diphosphates.…”

Section: Discussionmentioning

confidence: 99%

“…Approximately 6-15 min after infection, T7 RNA polymerase transcribes a second group of T7 genes, designated class II genes, that encode most of the proteins involved in DNA metabolism. The class II genes include those that encode the helicase/primase (gene 4), the DNA polymerase (gene 5) (1), and a single-stranded DNA-binding protein (gene 2.5) (2). A number of class II genes are not essential under laboratory growth conditions and have not yet been assigned specific functions (1).…”

mentioning

confidence: 99%

Gene 1.7 of bacteriophage T7 confers sensitivity of phage growth to dideoxythymidine

Tran

Rezende

Qimron

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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Bacteriophage T7 DNA polymerase efficiently incorporates dideoxynucleotides into DNA, resulting in chain termination. Dideoxythymidine (ddT) present in the medium at levels not toxic to Escherichia coli inhibits phage T7. We isolated 95 T7 phage mutants that were resistant to ddT. All contained a mutation in T7 gene 1.7, a nonessential gene of unknown function. When gene 1.7 was expressed from a plasmid, T7 phage resistant to ddT still arose; analysis of 36 of these mutants revealed that all had a single mutation in gene 5, which encodes T7 DNA polymerase. This mutation changes tyrosine-526 to phenylalanine, which is known to increase dramatically the ability of T7 DNA polymerase to discriminate against dideoxynucleotides. DNA synthesis in cells infected with wild-type T7 phage was inhibited by ddT, suggesting that it resulted in chain termination of DNA synthesis in the presence of gene 1.7 protein. Overexpression of gene 1.7 from a plasmid rendered E. coli cells sensitive to ddT, indicating that no other T7 proteins are required to confer sensitivity to ddT.deoxynucleotide kinase ͉ dideoxynucleosides ͉ DNA metabolism ͉ T7 DNA polymerase ͉ thymidine kinase W hen bacteriophage T7 infects Escherichia coli, the host RNA polymerase transcribes T7 early genes, designated class I genes. This group includes the T7 RNA polymerase (gene 1) (1). Approximately 6-15 min after infection, T7 RNA polymerase transcribes a second group of T7 genes, designated class II genes, that encode most of the proteins involved in DNA metabolism. The class II genes include those that encode the helicase/primase (gene 4), the DNA polymerase (gene 5) (1), and a single-stranded DNA-binding protein (gene 2.5) (2). A number of class II genes are not essential under laboratory growth conditions and have not yet been assigned specific functions (1).Little is known about the process of deoxynucleotide metabolism in T7-infected E. coli. T7 derives most of the phosphorus incorporated into its DNA from the breakdown of E. coli DNA (3, 4). The host DNA is degraded to 5Ј-deoxynucleoside monophosphates (dNMPs) by the joint action of the gene 3 endonuclease (5) and gene 6 exonuclease (6). It is not known whether the phage encodes its own enzymes to convert dNMPs to the dNTP precursors for DNA synthesis. E. coli encodes four different dNMP kinases, each specific for one dNMP (7,8). At least one of these kinases, CMP kinase, encoded by cmk, is essential for T7 growth (9). In turn, nucleoside diphosphokinase is the enzyme primarily responsible for conversion of dNDPs to dNTPs (10). Phage T4 encodes its own set of dNMP kinases with specificities that differ from that of the E. coli enzymes, whereas it uses the E. coli nucleoside diphosphokinase to convert the dNDPs to dNTPs (11). Phage T4 also encodes a thymidine kinase (tdk) (12, 13). Neither E. coli nor phage T7 will readily use exogenous thymine for DNA synthesis; in contrast, phage T4 efficiently incorporates exogenous thymine into its DNA (13).One of the interesting properties of T7 DNA polymerase is it...

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“…Protein-The ssDNA-binding protein encoded by gene 2.5 (gp2.5) of bacteriophage T7 is essential for T7 growth (46). gp2.5 not only removes secondary structure in ssDNA; it also physically interacts with other T7 replication proteins to coordinate the multiple reactions occurring at the replication fork (7,36).…”

Section: Stimulation Of Gp6 Activity By T7 Gene 25 Ssdna-bindingmentioning

confidence: 99%

Flap Endonuclease Activity of Gene 6 Exonuclease of Bacteriophage T7

Mitsunobu

Zhu

Lee

et al. 2014

Journal of Biological Chemistry

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Background: Flap endonucleases remove 5Ј-single-stranded DNA termini. Results: T7 gene 6 exonuclease is a flap endonuclease that cleaves 5Ј-single-stranded termini one nucleotide into the duplex. Conclusion:The flap endonuclease activity catalyzes the removal 5Ј-single-stranded tails arising from duplex DNA. Significance: The specificity of gene 6 protein identifies it as a flap endonuclease that can remove unusual structures of recombination and replication.

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Bacteriophage T7 gene 2.5 protein: an essential protein for DNA replication.

Abstract: The product of gene 2.5 of bacteriophage T7, a single-stranded DNA binding protein, physically interacts with the phage-encoded gene S protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities.

Cited by 63 publications

References 49 publications

Acidic C-terminal tail of the ssDNA-binding protein of bacteriophage T7 and ssDNA compete for the same binding surface

Acidic C-terminal tail of the ssDNA-binding protein of bacteriophage T7 and ssDNA compete for the same binding surface

Gene 1.7 of bacteriophage T7 confers sensitivity of phage growth to dideoxythymidine

Flap Endonuclease Activity of Gene 6 Exonuclease of Bacteriophage T7

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