The plasma membrane is the defining feature of cells and serves as their interface with the external environment. Current models posit that it is laterally organized in domains of distinct protein and lipid composition that are important for the efficient orchestration of key reactions of transport and communication, for example involved in endocytosis. However, despite tremendous advances in the comprehension of membrane processes and structure, physiological functions of plasma membrane organization are not yet clear. To overcome limitations of complex models used for investigation, we will focus on studying plasma membrane organization in yeast. This model system has prominent protein and lipid segregation in the plasma membrane and is amenable to molecular, biochemical, genetic and systems approaches. In this project, we will capitalize on our discovery of eisosomes, large protein complexes underlying the plasma membrane, as principal organizers plasma membrane domains in yeast.
We aim to define the molecular mechanisms and cellular functions of plasma membrane organization. We hypothesize that eisosomes assemble into a stable membrane scaffold that plays fundamental roles in phosphoinositide cell biology and endocytosis. To test this model, we will use directed biochemical, structural biology, cell biology experiments combined with unbiased analytical tools, including state-of-the-art proteomics and systematic genetics. By tackling these central questions on plasma membrane biology and elucidating the function of membrane organization by eisosomes, we will address a fundamental cell biology problem. Salient features of biological systems, including the structure of eisosome proteins are most often evolutionary conserved. Our findings will therefore likely have a broad impact on membrane research. They might also have therapeutic implications for a wide range of human pathologies where plasma membrane organization is implicated.

Public Health Relevance

In the proposed project, we will determine the mechanism and physiological functions of plasma membrane organization by eisosomes, especially in phosphoinositides metabolism and endocytosis. By addressing these questions, we will reveal fundamental principles of plasma membrane biology with broad implications.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM095982-03
Application #
8593302
Study Section
Membrane Biology and Protein Processing (MBPP)
Program Officer
Ainsztein, Alexandra M
Project Start
2012-02-15
Project End
2015-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
3
Fiscal Year
2014
Total Cost
$284,715
Indirect Cost
$113,715
Name
Yale University
Department
Anatomy/Cell Biology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Fröhlich, Florian; Christiano, Romain; Olson, Daniel K et al. (2014) A role for eisosomes in maintenance of plasma membrane phosphoinositide levels. Mol Biol Cell 25:2797-806
Harrison, Megan S; Hung, Chia-Sui; Liu, Ting-ting et al. (2014) A mechanism for retromer endosomal coat complex assembly with cargo. Proc Natl Acad Sci U S A 111:267-72
Bazzini, Ariel A; Johnstone, Timothy G; Christiano, Romain et al. (2014) Identification of small ORFs in vertebrates using ribosome footprinting and evolutionary conservation. EMBO J 33:981-93