Title: | An amino terminal prosequence is required for efficient synthesis of S. cerevisiae a-factor |
Address: | "Department of Biochemistry, University of Iowa, Iowa City 52242, USA" |
DOI: | 10.1016/s0167-4889(96)00153-x |
ISSN/ISBN: | 0006-3002 (Print) 0006-3002 (Linking) |
Abstract: | "The Saccharomyces cerevisiae a-mating pheromones are 12 amino acid lipopeptides whose secretion is dependent on the ABC transporter, Ste6p. The pheromones are synthesized as 36 and 38 amino acid precursors that terminate in a CaaX box (C is Cys, a is an aliphatic residue, and X is the C-terminal amino acid). Posttranslational processing of the a-factor precursors includes at least 5 events. C-terminal processing of the CaaX box includes farnesylation of Cys, removal of the -aaX residues, and methylation of the cysteine alpha-carboxyl group. The N-terminal steps involve proteolytic cleavages that remove the prosequences. In this report, we have investigated the role of posttranslational modification in the generation of functional a-factor. Wild type, mutant and chimeric forms of a-factor have been expressed in yeast and assessed for their abilities to serve as sources of functional a-factor. We have found that although modification of the CaaX box is necessary, it is not sufficient to generate bioactive a-factor. The amino terminal prosequences are also required. Deletion of these sequences reduces intracellular levels of a-factor resulting in sterility. Glutathione-S-transferase (GST)-a-factor fusions undergo CaaX box processing and membrane localization, but are not substrates for the N-terminal proteases and fail to interact with Ste6p. These results suggest that the amino terminal precursor sequences play a direct role in the generation of functional a-factor" |
Keywords: | "ATP-Binding Cassette Transporters/metabolism Amino Acid Sequence Fungal Proteins/metabolism Gene Expression Glutathione Transferase/metabolism *Glycoproteins Lipoproteins/*biosynthesis/chemistry Methylation Microscopy, Confocal Molecular Sequence Data Phe;" |
Notes: | "MedlineQuinby, G E Deschenes, R J eng HL42385/HL/NHLBI NIH HHS/ Research Support, U.S. Gov't, P.H.S. Netherlands 1997/03/27 Biochim Biophys Acta. 1997 Mar 27; 1356(1):23-34. doi: 10.1016/s0167-4889(96)00153-x" |