Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues

Yeast CEN plasmids expressing ALP and RS-ALP were constructed as follows: a 4-kbp BamHl fragment from pAL145 containing the PH08 gene was subeloned into the BamHI site of pKS +. The resulting plasmid (pSN93) was used for oligonucleotide mutagenesis to replace sequences encoding amino acids 11-17 of ALP with sequences encoding residues 81-90 of DPAP A creating the plasmid pSN94. The resulting amino acid sequence is as follows: (NH2...EQRRESFQFNDIDS...) where the DPAP A sequence is underlined. BamHI fragments from pSN93 and pSN94 were subcloned into pRS316 (CEN, URA3 based vector) resulting in pSN92 and pSN97, respectively, and into pRS313 (CEN, HIS3 based vector) creating pSN124 and pSN125, respectively.