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

Jui-Cheng Hsieh, Seung K U Yoo, Junetsu Ito

Research output: Contribution to journalArticle

19 Citations (Scopus)

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 α-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.

Original languageEnglish (US)
Pages (from-to)8665-8669
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume87
Issue number21
StatePublished - 1990

Fingerprint

DNA Replication
Arginine
Proteins
Bacteriophage PRD1
Phenylglyoxal
Conserved Sequence
Mutant Proteins
DNA-Directed DNA Polymerase
Site-Directed Mutagenesis
Amino Acid Sequence
Plasmids
Phosphates
Binding Sites
Amino Acids
DNA
Genes

Keywords

  • Deoxynucleotide binding site
  • DNA-terminal protein
  • Site-specific mutagenesis

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

An essential arginine residue for initiation of protein-primed DNA replication. / Hsieh, Jui-Cheng; Yoo, Seung K U; Ito, Junetsu.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 87, No. 21, 1990, p. 8665-8669.

Research output: Contribution to journalArticle

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