Inter-species complementation of the translocon beta subunit requires only its transmembrane domain.
In eukaryotes, proteins enter the secretory pathway through the translocon pore of the endoplasmic reticulum. This protein translocation channel is composed of three major subunits, called Sec61alpha, beta and gamma in mammals. Unlike the other subunits, the beta subunit is dispensable for translocation and cell viability in all organisms studied. Intriguingly, the knockout of the Sec61beta encoding genes results in different phenotypes in different species. Nevertheless, the beta subunit shows a high level of sequence homology across species, suggesting the conservation of a biological function that remains ill-defined. To address its cellular roles, we characterized the homolog of Sec61beta in the fission yeast Schizosaccharomyces pombe (Sbh1p). Here, we show that the knockout of sbh1(+) results in severe cold sensitivity, increased sensitivity to cell-wall stress, and reduced protein secretion at 23 degrees C. Sec61beta homologs from Saccharomyces cerevisiae and human complement the knockout of sbh1(+) in S. pombe. As in S. cerevisiae, the transmembrane domain (TMD) of S. pombe Sec61beta is sufficient to complement the phenotypes resulting from the knockout of the entire encoding gene. Remarkably, the TMD of Sec61beta from S. cerevisiae and human also complement the gene knockouts in both yeasts. Together, these observations indicate that the TMD of Sec61beta exerts a cellular function that is conserved across species.
Leroux
Alexandre
A
Department of Biochemistry, Université de Montréal, Montréal, Québec, Canada.
Rokeach
Luis A
LA
D000595:Amino Acid Sequence; D017124:Conserved Sequence; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D005816:Genetic Complementation Test; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D000069816:SEC Translocation Channels; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D013045:Species Specificity
D005656:Fungal Proteins; D008565:Membrane Proteins; D000069816:SEC Translocation Channels