Mapping of the DNA linking tyrosine residue of the PRD1 terminal protein

Shih Yi Shiue, Jui-Cheng Hsieh, Junetsu Ito

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

DNA replication of PRD1, a lipid-containing phage, is initiated by a protein-priming mechanism. The terminal protein encoded by gene 8 acts as a protein primer in DNA synthesis by forming an initiation complex with the 5′-terminal nucleotide, dGMP. The linkage between the terminal protein and the 5′ terminal nucleotide is a tyrosylphosphodiester bond. The PRD1 terminal protein contains 13 tyrosine residues in a total of 259 amino acids. By site-directed mutagenesis of cloned PRD1 gene 8, we replaced 12 of the 13 tyrosine residues in the terminal protein with phenylalanine and the other tyrosine residue with asparagine. Functional analysis of these mutant terminal proteins suggested that tyrosine-190 is the linking amino acid that forms a covalent bond with dGMP. Cyanogen bromide cleavage studies also implicated tyrosine-190 as the DNA-linking amino acid residue of the PRD1 terminal protein. Our results further show that tyrosine residues at both the amino-terminal and the carboxyl-terminal regions are important for the initiation complex forming activity. Predicted secondary structures for the regions around the DNA linking amino acid residues were compared in three terminal proteins (φ29, adenovirus-2, and PRD1). While the linking amino acids serine-232 (φ29) and serine-577 (adenovirus-2) are found in β-turns in hydrophilic regions, the linking tyrosine-190 of the PRD1 terminal protein is found in a β-sheet in a hydrophobic region.

Original languageEnglish (US)
Pages (from-to)3805-3810
Number of pages6
JournalNucleic Acids Research
Volume19
Issue number14
StatePublished - Jul 25 1991

Fingerprint

Linking
Tyrosine
DNA
Proteins
Protein
Amino Acids
Amino acids
Serine
Nucleotides
Cyanogen Bromide
Genes
DNA Primers
Asparagine
Site-directed mutagenesis
Gene
Mutant Proteins
Site-Directed Mutagenesis
Phenylalanine
DNA Replication
Mutagenesis

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Mapping of the DNA linking tyrosine residue of the PRD1 terminal protein. / Shiue, Shih Yi; Hsieh, Jui-Cheng; Ito, Junetsu.

In: Nucleic Acids Research, Vol. 19, No. 14, 25.07.1991, p. 3805-3810.

Research output: Contribution to journalArticle

Shiue, Shih Yi ; Hsieh, Jui-Cheng ; Ito, Junetsu. / Mapping of the DNA linking tyrosine residue of the PRD1 terminal protein. In: Nucleic Acids Research. 1991 ; Vol. 19, No. 14. pp. 3805-3810.
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