Akr1p-dependent palmitoylation of Yck2p yeast casein kinase 1 is necessary and sufficient for plasma membrane targeting.
The Yck2 protein is a plasma membrane-associated casein kinase 1 isoform that attaches to membranes via palmitoylation of its C terminus. We have demonstrated that Yck2p traffics to the plasma membrane on secretory vesicles. Because Akr1p, the palmitoyl transferase for Yck2p, is located on Golgi membranes, it is likely that Yck2p first associates with Golgi membranes, and then is somehow recruited to budding plasma membrane-destined vesicles. We show here that residues 499-546 are sufficient for minimal Yck2p palmitoylation and plasma membrane localization. We previously described normal plasma membrane targeting of a Yck2p construct with the final five amino acids of Ras2p substituting for the final two Cys residues of Yck2p. This Yck2p variant no longer requires Akr1p for membrane association, but targets normally. We have generated the C-terminal deletions previously shown to affect Yck2p membrane association in this variant to determine which residues are important for targeting and/or modification. We find that all of the sequences previously identified as important for plasma membrane association are required only for Akr1p-dependent modification. Furthermore, palmitoylation is sufficient for specific association of Yck2p with secretory vesicles destined for the plasma membrane. Finally, both C-terminal Cys residues are palmitoylated, and dual acylation is required for efficient membrane association.
Babu
Praveen
P
Department of Biochemistry and Molecular Biology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA.
Deschenes
Robert J
RJ
Robinson
Lucy C
LC
D000217:Acyltransferases; D047389:Casein Kinase I; D047388:Casein Kinases; D020134:Catalytic Domain; D002462:Cell Membrane / Q000378:metabolism; D003545:Cysteine / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D007527:Isoenzymes; D008856:Microscopy, Fluorescence; D010169:Palmitic Acids / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017368:Protein Prenylation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D017384:Sequence Deletion; D014316:Tritium; D015003:Yeasts / Q000166:cytology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
D007527:Isoenzymes; D010169:Palmitic Acids; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014316:Tritium; D000217:Acyltransferases; C107846:AKR1 protein, S cerevisiae; D011494:Protein Kinases; D047389:Casein Kinase I; D047388:Casein Kinases; C479666:YCK2 protein, S cerevisiae; D003545:Cysteine