Modular Organization of T4 DNA Polymerase
Open accessAbstract
We describe the use of a phylogenetic approach to analyze the modular organization of the single-chained (898 amino acids) and multifunctional DNA polymerase of phage T4. We have identified, cloned in expression vectors, and sequenced the DNA polymerase gene (gene 43) of phage RB69, a distant relative of T4. The deduced primary structure of the RB69 protein (RB69 gp43) differs from that of T4 gp43 in discrete clusters of short sequence that are interspersed with clusters of high similarity between the two proteins. Despite these differences, the two enzymes can substitute for each other in phage DNA replication, although T4 gp43 does exhibit preference to its own genome. A 55-amino acid internal gp43 segment of high sequence divergence between T4 and RB69 could be replaced in RB69 gp43 with the corresponding segment from T4 without loss of replication function. The reciprocal chimera and a deletion mutant of the T4 gp43 segment were both inactive for replication and specifically inhibitory ("dominant lethal") to the T4 wild-type allele. The results show that phylogenetic markers can be used to construct chimeric and truncated froms of gp43 that, although inactive for replication, can still exhibit biological specificity.
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PI3 kinase inhibitor wortmannin inhibits insulin but not contraction-stimulated glucose uptake.
“1) PI3 kinase inhibitor wortmannin inhibits insulin but not contraction-stimulated glucose uptake. Contraction does not promote IRS-1 autophosphorylation or increase PI3 kinase activity [103-106].”
From AMPK and Exercise: Glucose Uptake and Insulin Sensitivity (2013) · cites this paper as [103]
Contraction does not promote IRS-1 autophosphorylation or increase PI3 kinase activity.
“1) PI3 kinase inhibitor wortmannin inhibits insulin but not contraction-stimulated glucose uptake. Contraction does not promote IRS-1 autophosphorylation or increase PI3 kinase activity [103-106].”
From AMPK and Exercise: Glucose Uptake and Insulin Sensitivity (2013) · cites this paper as [103]
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Genetic Insertions and Diversification of the PolB-Type DNA Polymerase (gp43) of T4-Related Phages
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Self-citationAnatomy of Glucose Transporters in Skeletal Muscle
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Evolution of RNA-binding Specificity in T4 DNA Polymerase
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Self-citationA New Look at Old Mutants of T4 DNA Polymerase
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The Role of the Phosphoinositide Pathway in Hormonal Regulation of the Epithelial Sodium Channel
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Diversity of structure and function of DNA polymerase (gp43) of T4-related bacteriophages
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Divergence of the mRNA targets for the Ssb proteins of bacteriophages T4 and RB69
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Self-citationProtein Determinants of RNA Binding by DNA Polymerase of the T4-related Bacteriophage RB69
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Glucose Metabolism in Pregnancy
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The bacteriophage T4 replisome: a model system for understanding DNA replication mechanisms
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Structural insights into the exchange mechanism of a replicative DNA polymerase
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Intein Reporter and Selection Systems
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- Authors
- Wang, C.-C., Yeh, L.-S., & Karam, J. D.
- Affiliations
- Tulane University (US)
- Corresponding author
- Karam, Jim D.
- ORCID
- Wang, Chien-Chia
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1995-11
- Volume
- 270
- Issue
- 44
- Pages
- 26558-26564
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W1970574123
- Cited by
- 94
- References
- 33
- Citations by year
- 2025: 2 · 2023: 1 · 2022: 1 · 2020: 1 · 2014: 2 · 2013: 3
- FWCI
- 10.92
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health (5r01gm018842-20)
- National Institute of General Medical Sciences (GM 18842)
- Topics
- Physical Sciences › Environmental Science › Ecology › Bacteriophages and microbial interactions
- Physical Sciences › Materials Science › Materials Chemistry › Enzyme Structure and Function
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- T4 DNA polymerase, protein domain organization, fusion protein, dominant lethals, sequence divergence, phage DNA replication, phylogenetic analysis, deletion mutants
- MeSH terms
- Amino Acid Sequence, Base Sequence, DNA-Directed DNA Polymerase, DNA, Viral, Genes, Viral, Molecular Sequence Data, Phylogeny, Recombinant Proteins, Sequence Homology, Nucleic Acid, Genetic Variation, Viral Proteins, Blotting, Southern, Polymerase Chain Reaction, Bacteriophage T4, Sequence Homology, Amino Acid
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