Identification of interdependent signals required for anterograde traffic of the ATP-binding cassette transporter protein Yor1p.
The plasma membrane ATP-binding cassette (ABC) transporter Yor1p mediates oligomycin resistance in Saccharomyces cerevisiae. Its protein sequence places it in the multidrug resistance protein/cystic fibrosis transmembrane conductance regulator subfamily of ABC transporters. A key regulatory step in the biogenesis of this family of ABC transporter proteins is at the level of transport from the endoplasmic reticulum (ER) on through the secretory pathway. To explore the protein sequence requirements for Yor1p to move from the ER to its site of function at the plasma membrane, a series of truncation and alanine replacement mutations were constructed in Yor1p. This analysis detected two sequence motifs similar to the DXE element that has recently been found in other proteins that exit the ER. Loss of the N-terminal DXE element eliminated function of the protein, whereas loss of the C-terminal element only slightly reduced function of the resulting mutant Yor1p. Strikingly, although both of the single mutant proteins were stable, production of the double mutant caused dramatic destabilization of Yor1p. These data suggest that this large polytopic membrane protein requires multiple signals for normal forward trafficking, and elimination of this information may cause the mutant protein to be transferred to a degradative fate.
Epping
Eric A
EA
Department of Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242, USA.
Moye-Rowley
W Scott
WS
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000595:Amino Acid Sequence; D001483:Base Sequence; D017931:DNA Primers; D004721:Endoplasmic Reticulum / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006868:Hydrolysis; D008164:Luminescent Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction / Q000502:physiology*
D018528:ATP-Binding Cassette Transporters; D017931:DNA Primers; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; C097077:YOR1 protein, S cerevisiae; D049452:Green Fluorescent Proteins