Physical monitoring of mating type switching in Saccharomyces cerevisiae

Bernadette Connolly, C I White, J E Haber

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

The kinetics of mating type switching in Saccharomyces cerevisiae can be followed at the DNA level by using a galactose-inducible HO (GAL-HO) gene to initiate the event in synchronously growing cells. From the time that HO endonuclease cleaves MAT a until the detection of MAT alpha DNA took 60 min. When unbudded G1-phase cells were induced, switched to the opposite mating type in "pairs." In the presence of the DNA synthesis inhibitor hydroxyurea, HO-induced cleavage occurred but cells failed to complete switching. In these blocked cells, the HO-cut ends of MATa remained stable for at least 3 h. Upon removal of hydroxyurea, the cells completed the switch in approximately 1 h. The same kinetics of MAT switching were also seen in asynchronous cultures and when synchronously growing cells were induced at different times of the cell cycle. Thus, the only restriction that confined normal homothallic switching to the G1 phase of the cell cycle was the expression of HO endonuclease. Further evidence that galactose-induced cells can switch in the G2 phase of the cell cycle was the observation that these cells did not always switch in pairs. This suggests that two chromatids, both cleaved with HO endonuclease, can interact independently with the donors HML alpha and HMRa.
Original languageEnglish
Pages (from-to)2342-2349
Number of pages8
JournalMolecular and Cellular Biology
Volume8
Issue number6
Publication statusPublished - 1 Jun 1988

    Fingerprint

Keywords

  • DNA
  • DNA Restriction Enzymes
  • Electrophoresis, Agar Gel
  • Enzyme Induction
  • Galactose
  • Genes, Fungal
  • Genes, Mating Type, Fungal
  • Kinetics
  • Peptides
  • Promoter Regions, Genetic
  • Saccharomyces cerevisiae
  • Time Factors

Cite this