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. 2000 Jul 10;150(1):77–88. doi: 10.1083/jcb.150.1.77

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© 2000 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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A, per5-1 is defective in N-linked glycosylation. Wild-type (W303a) and per5-1 cells were metabolically pulse-labeled for 5 min and chased for 0, 15, and 30 min. CPY was immunoprecipitated from detergent lysates and divided into two aliquots for each time point. One aliquot was treated with endoglycosidase H and the other was mock-treated. Proteins were separated on a 10% polyacrylamide gel and visualized by fluorography. The positions of the different pro and mature glycoforms of CPY are indicated as described in Fig. 2. Endo H deglycosylated pro (dg-proCPY) and mature forms (dg-mCPY) are indicated to the right. B, per5-1 is an allele of RFT1. Wild-type, per5-1, and per5-1 (pDN387) cells were pulse-labeled for 5 min and chased for 0 and 30 min. pDN387 is a centromeric vector containing the RFT1 gene. Immunoprecipitates of Gas1p were resolved by a 10–15% polyacrylamide gradient gel and visualized by autoradiography. The normal ER and mature (Golgi/PM) forms are shown.