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Journal of Biosciences

, Volume 36, Issue 1, pp 79–96 | Cite as

Requirements of Slm proteins for proper eisosome organization, endocytic trafficking and recycling in the yeast Saccharomyces cerevisiae

  • Chitra Kamble
  • Sandhya Jain
  • Erin Murphy
  • Kyoungtae KimEmail author
Article

Abstract

Eisosomes are large immobile assemblies at the cortex of a cell under the membrane compartment of Can1 (MCC) in yeast. Slm1 has recently been identified as an MCC component that acts downstream of Mss4 in a pathway that regulates actin cytoskeleton organization in response to stress. In this study, we showed that inactivation of Slm proteins disrupts proper localization of the primary eisosome marker Pil1, providing evidence that Slm proteins play a role in eisosome organization. Furthermore, we found that slm ts mutant cells exhibit actin defects in both the ability to polarize cortical F-actin and the formation of cytoplasmic actin cables even at the permissive temperature (30°C). We further demonstrated that the actin defect accounts for the slow traffic of FM4-64-labelled endosome in the cytoplasm, supporting the notion that intact actin is essential for endosome trafficking. However, our real-time microscopic analysis of Abp1-RFP revealed that the actin defect in slm ts cells was not accompanied by a noticeable defect in actin patch internalization during receptor-mediated endocytosis. In addition, we found that slm ts cells displayed impaired membrane recycling and that recycling occurred in an actin-independent manner. Our data provide evidence for the requirement of Slm proteins in eisosome organization and endosome trafficking and recycling.

Keywords

Actin Actin patch Eisosome Endocytosis Endosome FM4-64 Pil1 Recycling Slm1/2 Trafficking 

Abbreviations

FITC

fluorescein isothiocyanate

LCB

long-chain bases

LY

Lucifer yellow

MCC

membrane compartment of Can1

MCP

membrane compartment of Pma1

NA

numerical aperture

PH

pleckstrin homology

RME

receptor-mediated endocytosis

YPD

yeast peptone dextrose

Notes

Acknowledgements

We are grateful to Scott Emr for providing slm ts mutant strains and Mark Longtine for providing the PCR templates. We thank Paul Durham and Mark Richter for providing unrestricted access to laboratories and equipment. This work was supported by a National Scientific Foundation Grant 0923024 (to KK) and by thesis funding from Missouri State University (to CK).

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Copyright information

© Indian Academy of Sciences 2011

Authors and Affiliations

  • Chitra Kamble
    • 1
  • Sandhya Jain
    • 1
  • Erin Murphy
    • 1
  • Kyoungtae Kim
    • 1
    Email author
  1. 1.Department of BiologyMissouri State UniversitySpringfieldUSA

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