Sequencing and enhanced expression of the gene encoding diadenosine 5′,5‴-P1, P4-tetraphosphate (Ap4A) phosphorylase in Saccharomyces cerevisiae

Varsha Kaushal, Diana M. Avila, Stephen C. Hardies, Larry D. Barnes

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The gene, DTP, coding for diadenosine 5′, 5‴-P1, P4-tetraphosphate (Ap4P) phosphorylase was isolated from a Saccharomyces cerevisiae genomic DNA library in λgt11. In yeast and Escherichia coli transformed with the multicopy vector, YEp352, containing the cloned DTP gene, the Ap4A phosphorylase was produced at levels nine- to 17-fold higher than in untransformed hosts. The nucleotide (nt) sequence was determined. The gene codes for a polypeptide chain of 321 amino acids (aa). Two-aa sequence motifs of possible significance were identified: a potential adenine nt binding site and a potential phosphorylation site. The DTP gene is located on yeast chromosome III and is present as a single copy. Although multicopy vector expression increased the Ap4A phosphorylase activity ninefold above the endogenous activity in transformed yeast, the intracellular concentration of Ap4A did not decrease and the growth rate of the yeast was unchanged.

Original languageEnglish (US)
Pages (from-to)79-84
Number of pages6
JournalGene
Volume95
Issue number1
DOIs
StatePublished - Oct 30 1990

Fingerprint

Phosphorylases
Saccharomyces cerevisiae
Yeasts
Gene Expression
Genes
Amino Acid Motifs
Adenine Nucleotides
Genomic Library
Gene Library
Amino Acid Sequence
Chromosomes
Binding Sites
Phosphorylation
Escherichia coli
Amino Acids
Peptides
diadenosine tetraphosphate
Growth

Keywords

  • amplification
  • chromosome hybridization
  • Escherichia coli
  • growth rate
  • multicopy vector
  • phage λgt11
  • Recombinant DNA
  • yeast

ASJC Scopus subject areas

  • Genetics

Cite this

Sequencing and enhanced expression of the gene encoding diadenosine 5′,5‴-P1, P4-tetraphosphate (Ap4A) phosphorylase in Saccharomyces cerevisiae. / Kaushal, Varsha; Avila, Diana M.; Hardies, Stephen C.; Barnes, Larry D.

In: Gene, Vol. 95, No. 1, 30.10.1990, p. 79-84.

Research output: Contribution to journalArticle

Kaushal, Varsha ; Avila, Diana M. ; Hardies, Stephen C. ; Barnes, Larry D. / Sequencing and enhanced expression of the gene encoding diadenosine 5′,5‴-P1, P4-tetraphosphate (Ap4A) phosphorylase in Saccharomyces cerevisiae. In: Gene. 1990 ; Vol. 95, No. 1. pp. 79-84.
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