Active-site mutations in the Xrn1p exoribonuclease of Saccharomyces cerevisiae reveal a specific role in meiosis

Jachen A. Solinger, Donatella Pascolini, Wolf Dietrich Heyer

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Xrn1p of Saccharomyces cerevisiae is a major cytoplasmic RNA turnover exonuclease which is evolutionarily conserved from yeasts to mammals. Deletion of the XRN1 gene causes pleiotropic phenotypes, which have been interpreted as indirect consequences of the RNA turnover defect. By sequence comparisons, we have identified three loosely defined, common 5'-3' exonuclease motifs. The significance of motif II has been confirmed by mutant analysis with Xrn1p. The amino acid changes D206A and D208A abolish singly or in combination the exonuclease activity in vivo. These mutations show separation of function. They cause identical phenotypes to that of xrn1Δ in vegetative cells but do not exhibit the severe meiotic arrest and the spore lethality phenotype typical for the deletion. In addition, xrn1-D208A does not cause the severe reduction in meiotic popout recombination in a double mutant with dmc1 as does xrn1Δ. Biochemical analysis of the DNA binding, exonuclease, and homologous pairing activity of purified mutant enzyme demonstrated the specific loss of exonuclease activity. However, the mutant enzyme is competent to promote in vitro assembly of tubulin into microtubules. These results define a separable and specific function of Xrn1p in meiosis which appears unrelated to its RNA turnover function in vegetative cells.

Original languageEnglish (US)
Pages (from-to)5930-5942
Number of pages13
JournalMolecular and Cellular Biology
Volume19
Issue number9
StatePublished - Sep 1999

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Exoribonucleases
Exonucleases
Meiosis
Saccharomyces cerevisiae
Catalytic Domain
Mutation
RNA
Phenotype
Exodeoxyribonucleases
Genetic Pleiotropy
Gene Deletion
Enzymes
Tubulin
Spores
Microtubules
Genetic Recombination
Mammals
Yeasts
Amino Acids

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

Active-site mutations in the Xrn1p exoribonuclease of Saccharomyces cerevisiae reveal a specific role in meiosis. / Solinger, Jachen A.; Pascolini, Donatella; Heyer, Wolf Dietrich.

In: Molecular and Cellular Biology, Vol. 19, No. 9, 09.1999, p. 5930-5942.

Research output: Contribution to journalArticle

Solinger, Jachen A. ; Pascolini, Donatella ; Heyer, Wolf Dietrich. / Active-site mutations in the Xrn1p exoribonuclease of Saccharomyces cerevisiae reveal a specific role in meiosis. In: Molecular and Cellular Biology. 1999 ; Vol. 19, No. 9. pp. 5930-5942.
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