This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Beclin 1 is a mammalian autophagy protein which was shown to play an important role in development, tumor suppression, neurodegeneration and cell death. Beclin 1 forms a complex with Vps34/phosphatidylinositol (PtdIns) 3-kinase (class III PI-3K), which mediates multiple vesicle trafficking pathways including endocytosis and autophagy. However, the precise role of Beclin 1-Vps34/PtdIns3K complex in the regulation of autophagy remains to be elucidated. Through a study that combines mouse genetics and biochemistry, we uncover several protein components that associate specifically with Beclin 1 complexes in mouse tissues. The biochemical analysis reveals a large in vivo Beclin 1 complex containing the known proteins Vps34/PtdIns3K, p150/Vps15 and UVRAG, as well as novel proteins BISC and BIRC. Characterization of the novel proteins suggests that BISC and BIRC modulate Vps34/PtdIns3K activity in opposite manner. This opposing effect of BISC and BIRC on the lipid kinase activity is correlated with their distinct regulation of autophagy: BISC positively regulates autophagy, whereas BIRC is involved in negative control of autophagy. Moreover, we find that Beclin 1 and BISC synergistically promote formation of double membrane vacuoles which are associated with Atg5/Atg12, while forced expression of BIRC results in aberrant late endosomal/lysosomal structures (independent of Beclin 1) and impaired autophagosome maturation. Our study suggests that, by forming distinct protein complexes, Beclin 1 and its binding proteins orchestrate the precise function of Vps34/PtdIns3K in regulating autophagy at multiple steps. A manuscript describing this work has been published (18. Y. Zhong, Q.J. Wang, X. Li, B.T. Chait, N. Heintz, Z. Yue """"""""Distinct regulation of autophagic activity by Atg14L and Rubicon associated with Beclin 1-phosphatidylinositol-3-kinase complex"""""""" Nature Cell Biology. 11 (2009) 468-76.)

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-38
Application #
8361544
Study Section
Special Emphasis Panel (ZRG1-BCMB-Q (40))
Project Start
2011-03-01
Project End
2012-03-31
Budget Start
2011-03-01
Budget End
2012-03-31
Support Year
38
Fiscal Year
2011
Total Cost
$1,304
Indirect Cost
Name
Rockefeller University
Department
Miscellaneous
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065
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Indiani, Chiara; O'Donnell, Mike (2013) A proposal: Source of single strand DNA that elicits the SOS response. Front Biosci (Landmark Ed) 18:312-23

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