Suppression of Ycf1p function by Cka1p-dependent phosphorylation is attenuated in response to salt stress

Kerry A. Pickin, Nkiruka Ezenwajiaku, Holly Overcash, Manish Sethi, Marc Knecht, Christian M. Paumi

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

The yeast vacuolar membrane protein Ycf1p and its mammalian counterpart, MRP1, belong to the ABCC subfamily of ATP-binding cassette transporters. Genetic evidence suggests that the yeast casein kinase 2α, Cka1p, negatively regulates Ycf1p function via phosphorylation of Ser251 within the N-terminus. In this study, we provide strong evidence that Cka1p regulates Ycf1p function via phosphorylation of Ser251. We show that the CK2 holoenzyme interacts with Ycf1p. However, genetic analysis suggests that only Cka1p is required for Ser251 phosphorylation, as the deletion of CKA1 significantly reduces Ser251 phosphorylation in vivo. Furthermore, purified recombinant Cka1p phosphorylates a Ycf1p-derived peptide containing Ser251. We also demonstrate that Ycf1p function is induced in response to high salt stress. Induction of the Ycf1p function strongly correlates with reduced phosphorylation of Ser251. Importantly, Cka1p activity in vivo is similarly reduced in response to salt stress, consistent with our finding that Cka1p directly phosphorylates Ser251 of Ycf1p. We provide genetic and biochemical evidence that strongly suggests that the induction of Ycf1p function is the result of decreased phosphorylation of Ser251. In conclusion, our work demonstrates a novel biochemical role for Cka1p regulation of Ycf1p function in the cellular response of yeast to salt stress.

Original languageEnglish
Pages (from-to)839-857
Number of pages19
JournalFEMS Yeast Research
Volume10
Issue number7
DOIs
StatePublished - Nov 1 2010
Externally publishedYes

Fingerprint

Salts
Phosphorylation
Yeasts
Casein Kinase II
Holoenzymes
ATP-Binding Cassette Transporters
Molecular Biology
Membrane Proteins
Peptides

Keywords

  • ABC transporter
  • Casein kinase 2
  • Cka1p
  • Saccharomyces cerevisiae
  • Salt stress
  • Ycf1p

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Microbiology

Cite this

Suppression of Ycf1p function by Cka1p-dependent phosphorylation is attenuated in response to salt stress. / Pickin, Kerry A.; Ezenwajiaku, Nkiruka; Overcash, Holly; Sethi, Manish; Knecht, Marc; Paumi, Christian M.

In: FEMS Yeast Research, Vol. 10, No. 7, 01.11.2010, p. 839-857.

Research output: Contribution to journalArticle

Pickin, Kerry A. ; Ezenwajiaku, Nkiruka ; Overcash, Holly ; Sethi, Manish ; Knecht, Marc ; Paumi, Christian M. / Suppression of Ycf1p function by Cka1p-dependent phosphorylation is attenuated in response to salt stress. In: FEMS Yeast Research. 2010 ; Vol. 10, No. 7. pp. 839-857.
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AU - Knecht, Marc

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AB - The yeast vacuolar membrane protein Ycf1p and its mammalian counterpart, MRP1, belong to the ABCC subfamily of ATP-binding cassette transporters. Genetic evidence suggests that the yeast casein kinase 2α, Cka1p, negatively regulates Ycf1p function via phosphorylation of Ser251 within the N-terminus. In this study, we provide strong evidence that Cka1p regulates Ycf1p function via phosphorylation of Ser251. We show that the CK2 holoenzyme interacts with Ycf1p. However, genetic analysis suggests that only Cka1p is required for Ser251 phosphorylation, as the deletion of CKA1 significantly reduces Ser251 phosphorylation in vivo. Furthermore, purified recombinant Cka1p phosphorylates a Ycf1p-derived peptide containing Ser251. We also demonstrate that Ycf1p function is induced in response to high salt stress. Induction of the Ycf1p function strongly correlates with reduced phosphorylation of Ser251. Importantly, Cka1p activity in vivo is similarly reduced in response to salt stress, consistent with our finding that Cka1p directly phosphorylates Ser251 of Ycf1p. We provide genetic and biochemical evidence that strongly suggests that the induction of Ycf1p function is the result of decreased phosphorylation of Ser251. In conclusion, our work demonstrates a novel biochemical role for Cka1p regulation of Ycf1p function in the cellular response of yeast to salt stress.

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