Molecular and Cellular Biochemistry

, Volume 346, Issue 1–2, pp 187–195 | Cite as

env1 Mutant of VPS35 gene exhibits unique protein localization and processing phenotype at Golgi and lysosomal vacuole in Saccharomyces cerevisiae

  • Editte Gharakhanian
  • Onyinyechi Chima-Okereke
  • Daniel K. Olson
  • Christopher Frost
  • M. Kathleen Takahashi


The yeast vacuole is functionally and structurally equivalent to the mammalian lysosome. Delivery of resident and cargo proteins to the lysosome is vital for proper cellular operations, and failure to correctly target proteins to the organelle is correlated with the development of neurodegenerative and lysosomal storage diseases. We previously reported a novel mutant screen for vacuolar trafficking defects in yeast Saccharomyces cerevisiae that resulted in the isolation of env1, an allelic mutant of VPS35. As a member of the retromer complex, Vps35p binds directly to cargos and facilitates their retrograde transport to trans Golgi from endosomes. Our previous studies established that env1 exhibits unique pleiotropic phenotype in comparison to other tested VPS35 alleles including severe growth sensitivity to hygromycin B and internal accumulation of the precursor form of the vacuolar enzyme carboxypeptidase Y. Here, through a combination of sub-cellular fractionation and indirect immunofluorescence microscopy, we confirm and extend the unique phenotype of env1 to processing and localization of additional proteins within the vacuolar trafficking pathway. In comparative studies with a null and an allelic mutant of VPS35, env1 exhibited unique processing defects of retromer-independent vacuolar membrane enzyme alkaline phosphatase at the vacuole and significant Golgi localization of retromer cargos Vps10p and Kex2p despite compromised trafficking at the Golgi and late endosome interface.


Yeast vacuole Lysosome Late endosome VPS35 Vesicular trafficking 


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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Editte Gharakhanian
    • 1
  • Onyinyechi Chima-Okereke
    • 1
  • Daniel K. Olson
    • 1
  • Christopher Frost
    • 1
  • M. Kathleen Takahashi
    • 1
    • 2
  1. 1.Department of Biological SciencesCalifornia State University at Long BeachLong BeachUSA
  2. 2.Biology DepartmentSanta Ana CollegeSanta AnaUSA

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