A mutation in the secretion pathway of the yeast Yarrowia lipolytica that displays synthetic lethality in combination with a mutation affecting the signal recognition particle.
In an attempt to identify proteins involved in the translocation step of protein secretion, a genetic screen was carried out in the yeast Yarrowia lipolytica. A conditional lethal mutant which has a defect in the 7S RNA of the signal recognition particle was mutagenized and screened for second-site mutations that specifically exacerbate its temperature sensitivity. This approach had previously allowed the characterization of an endoplasmic reticulum component, Sls1p, involved in protein translocation. A second mutation, sls2-1, was isolated that causes synthetic lethality when combined with the 7S RNA mutation. On its own, the sls2-1 mutation confers a temperature-sensitive growth phenotype. The secretory phenotype of the sls2 mutant consists in abnormal secretion of several polypeptides, and thus differs from the defect in secretory protein synthesis associated with the 7S RNA and sls1-1 mutations. Two new Y. lipolytica genes were identified which can relieve the growth defect of sls2-1 cells: SLS2 itself and SSL2, a multicopy suppressor of the temperature sensitivity of the sls2 mutant. The SLS2 gene encodes a polypeptide that can potentially be farnesylated and phosphorylated, and shares some homology with an S. cerevisiae protein of unknown function. Ssl2p resembles calmodulin-dependent serine/threonine protein kinases. These two proteins may interact to regulate protein sorting.
Boisramé
A
A
Laboratoire de Génetique moléculaire et cellulaire INRA, CNRS, Institut National Agronomique Paris-Grignon, Thiverval-Grignon, France. boisrame@cardere.grignon.inra.fr
Beckerich
J M
JM
Gaillardin
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004265:DNA Helicases / Q000235:genetics; D004260:DNA Repair; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics*; D005800:Genes, Fungal; D005804:Genes, Lethal; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016366:Open Reading Frames; D012331:RNA, Fungal / Q000235:genetics; D015183:Restriction Mapping; D004718:Saccharomycetales / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D018271:Signal Recognition Particle / Q000235:genetics*; D014466:Ultraviolet Rays
D005656:Fungal Proteins; D012331:RNA, Fungal; D018271:Signal Recognition Particle; C120217:Sls2 protein, Yarrowia lipolytica; D004265:DNA Helicases