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PMID: 2236078 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, P.H.S.

An essential arginine residue for initiation of protein-primed DNA replication.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 87 ·No. 21 ·1990-11-00 ·Pages 8665-9

Hsieh JC, Yoo SK, Ito J

Abstract

A group of proteins that act as primers for initiation of linear DNA replication are called DNA-terminal proteins (terminal proteins). We have found a short stretch of conserved amino acid sequence among the terminal proteins from six different sources. The location of this sequence motif is also similar among the different terminal-proteins. To determine the functional role of this terminal-protein domain in DNA replication, we have studied the bacteriophage PRD1 system. The PRD1 terminal protein and DNA polymerase genes were cloned into expression vectors, and the recombinant plasmids were used for constructing PRD1 terminal protein mutants. Site-directed mutagenesis and functional analysis showed that one of the two arginines (Arg-174) in the conserved sequence is critical for the initiation complex-forming activity of the PRD1 terminal protein. Replacement of Arg-174 by noncharged amino acids resulted in nonfunctional terminal protein. Phenylglyoxal, an alpha-dicarbonyl compound that reacts with the guanidino group of arginine, inhibits initiation complex formation between PRD1 terminal protein and dGMP. On the basis of these results, we propose that Arg-174 represents, at least in part, the binding site for phosphate groups of dGTP.

MeSH Terms
Amino Acid Sequence Arginine Base Sequence Binding Sites Coliphages/genetics DNA Replication Deoxyguanine Nucleotides/metabolism Escherichia coli/genetics Genes, Viral Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Nucleic Acid Conformation Protein Conformation Sequence Homology, Nucleic Acid Viral Proteins/genetics,metabolism Viral Structural Proteins/genetics
Chemicals
DNA terminal protein, Enterobacteria phage PRD1 Deoxyguanine Nucleotides Viral Proteins Viral Structural Proteins 2'-deoxyguanosine 5'-phosphate Arginine
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Hsieh J C
Department of Microbiology and Immunology, College of Medicine, University of Arizona, Tucson 85724.
Yoo S K
Ito J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.87.21.8665
Published
1990-11-00
Pages
8665-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55018
Subset
IM
Grants
NIGMS NIH HHS · GM28013 · United States
External Links
PubMed source DOI: 10.1073/pnas.87.21.8665 PMC Full Text View journal details
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