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. We are studying the interacting binding partners of two Jumonji domain containing proteins, JHD2 and ECM5. The first IPs using commercially available protein A tagged strains failed to lead to any interacting partners. The amount of yeast cells used was increased to 30 g per IP, as both proteins have low copy/cell numbers. In addition, different IP buffers were tried. Nevertheless, the IPs did not lead any interacting partners. Subsequently, we tagged JHD2 and ECM5 ourselves using the Rout lab protein A tag. ECM5 IP now resulted in SNT2 and RPD3 as interacting partners. SNT2 and RPD3 were in turn tagged with the Rout lab protein A tag and the reciprocal IPs confirmed the presence of SNT2 and ECM5 in the RPD3 IP and the presence of ECM5 and RPD3 in the SNT2 IP. The JHD2 IP was carried out testing 3 different IP conditions. Silver stain and Western blot data confirmed isolation of JHD2 in all 3 IPs, but the LC-MS analysis of the complex is still pending. PTM analysis of ECM5 and SNT2 are planned for the future.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000862-38
Application #
8361527
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
$2,607
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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