Multicopy STS1 restores both protein transport and ribosomal RNA stability in a new yeast sec23 mutant allele.
A thermosensitive mutant was selected on the basis of its resistance after a heat shock to the lethal effects of lomofungin, a drug that inhibits RNA synthesis. We demonstrate that the single mutation conferring thermosensitivity and lomofungin resistance is heteroallelic to sec23-1. This new allele of sec23 is designated sec23-11. The yeast SEC23 gene has previously been reported to function in protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. After a 1-min shift to elevated temperature, a sec23-1 mutant becomes defective in vesicle formation from the ER. We show here that upon a shift to high temperature, both sec23 alleles confer defects in ER-to-Golgi transport, and both mutants stop accumulating nascent RNA after 20 to 30 min. Transcription and early maturation of ribosomal RNA, the major RNA species, are normal in a sec23-11 mutant, but the matured ribosomal RNA is then degraded. This ribosomal RNA instability may reflect improper assembly of ribosomal subunits, due to failure to localize essential factors such as ribosomal proteins to the nucleus. In addition to the sec23 mutants, other ER-to-Golgi secretion mutants exhibit a strong RNA biosynthetic defect, and we conclude that continued post-ER protein transport, not just a functional SEC23 gene product (Sec23p), is required for ribosomal RNA stability. We also identified an extragenic multicopy suppressor of the sec23-11 mutant, the essential STS1 gene ("Sec Twenty-three Suppressor 1"). Besides restoring partial temperature resistance to the sec23-11 mutant, overexpression of STS1 also overcomes both the RNA synthesis and the protein transport blocks in this mutant. In contrast, multicopy STS1 exerts only small effects on the sec23-1 mutant. The predicted STS1 gene product (Sts1p) is a low-abundance protein of 36,500 daltons, with no signal peptide and no hydrophobic stretch of sufficient length to span a membrane. Like Sec23p, Sts1p is found in a cytosolic compartment. These features suggest that Sts1p may play a catalytic or regulatory role in conjunction with the more abundant Sec23p.
Liang
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S
Centre de Génétique Moléculaire, Centre National de la Recherche Scientifique, Gif-sur-Yvette/France.
Lacroute
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Képès
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D018528:ATP-Binding Cassette Transporters / Q000235:genetics*; D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport / Q000502:physiology; D004721:Endoplasmic Reticulum / Q000502:physiology / Q000648:ultrastructure; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism*; D005800:Genes, Fungal; D016153:Genes, Suppressor; D006056:Golgi Apparatus / Q000502:physiology / Q000648:ultrastructure; D008969:Molecular Sequence Data; D009154:Mutation; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012335:RNA, Ribosomal / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins
D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D012331:RNA, Fungal; D012335:RNA, Ribosomal; C085810:STS1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins