Analysis of Sec22p in endoplasmic reticulum/Golgi transport reveals cellular redundancy in SNARE protein function.
Membrane-bound soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins form heteromeric complexes that are required for intracellular membrane fusion and are proposed to encode compartmental specificity. In yeast, the R-SNARE protein Sec22p acts in transport between the endoplasmic reticulum (ER) and Golgi compartments but is not essential for cell growth. Other SNARE proteins that function in association with Sec22p (i.e., Sed5p, Bos1p, and Bet1p) are essential, leading us to question how transport through the early secretory pathway is sustained in the absence of Sec22p. In wild-type strains, we show that Sec22p is directly required for fusion of ER-derived vesicles with Golgi acceptor membranes. In sec22Delta strains, Ykt6p, a related R-SNARE protein that operates in later stages of the secretory pathway, is up-regulated and functionally substitutes for Sec22p. In vivo combination of the sec22Delta mutation with a conditional ykt6-1 allele results in lethality, consistent with a redundant mechanism. Our data indicate that the requirements for specific SNARE proteins in intracellular membrane fusion are less stringent than appreciated and suggest that combinatorial mechanisms using both upstream-targeting elements and SNARE proteins are required to maintain an essential level of compartmental organization.
Liu
Yiting
Y
Department of Biochemistry, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Barlowe
Charles
C
D001692:Biological Transport; D002474:Cell-Free System; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D015151:Immunoblotting; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000378:metabolism* / Q000502:physiology; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D021381:Protein Transport; D050683:R-SNARE Proteins; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000502:physiology; D050600:SNARE Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000502:physiology; D033921:Vesicular Transport Proteins
D008565:Membrane Proteins; D050683:R-SNARE Proteins; D011956:Receptors, Cell Surface; D050600:SNARE Proteins; D029701:Saccharomyces cerevisiae Proteins; C106909:Sec22 protein, S cerevisiae; D033921:Vesicular Transport Proteins; C107127:YKT6 protein, S cerevisiae