Michael P. Myers - US grants

[unreadable] DESCRIPTION (provided by applicant): We are requesting funding for a Thermo Finnigan LCQdecaXP+ ion trap mass spectrometer. Characterization of multi-protein complexes has become one of the most powerful methods for elucidating protein/gene function and for understanding how normal cellular functions, as well as, pathological conditions, such as cancer or diabetes, are controlled by these multi-protein complexes. This type of analysis provides an important foundation for understanding gene function and has become a critical component of research at Cold Spring Harbor Laboratory. In recent years, mass spectrometry has superceded other techniques for protein identification and characterization. This is largely due to the enhanced sensitivity, through put and flexibility provided by mass spectrometry. The Protein Chemistry Shared Resource in the Cold Spring Harbor Laboratory Cancer Center has become increasingly reliant on mass spectrometry and is requesting funding for a Thermo Finnigan LCQdeca XP+ mass spectrometer. The LCQdeca XP+ is a state of the art ion trap mass spectrometer, which is renowned for its sensitivity and analytical capabilities. The new instrumentation will enhance the sensitivity, reliability, and through put of this critical application and will have a major impact on the shared users of this grant and the larger Cold Spring Harbor community. This instrument will be incorporated into an existing mass spectrometry based protein chemistry shared resource, and will therefore be integrated into an existing organization that will oversee the management, operation, and maintenance of the new instrument. [unreadable] [unreadable]

DESCRIPTION (provided by applicant): The overall goal of this project is to understand the function of HCF proteins in cell division and proliferatior. The founding member of this family is the human herpes simplex virus (HSV) host cell factor HCF-1. HCF(also known as Cl, VCAF, and CFF) is an abundant chromatin-associated protein that is highly conserved i animals. It is a large protein that undergoes an unusual maturation process involving proteolysis that results i N- and C-terminal HCF-1N and HCF-1c polypeptides that remain stably associated with one another. Dunn HSV infection, HCF-1 plays an important regulatory role in the transcription of HSV immediate-early (IE genes through its association with the HSV transactivator VP16. In uninfected cells, HCF-1 plays a critica role in cell division and proliferation through its association with chromatin. HCF- 1 does not association directly with DNA but associates with chromatin through a defined protein-protein interaction module, Keich domain, also used to tether HCF-1 to the HSV genome through its interaction with VP 16. HCF-1 and related human protein HCF-2 share a single conserved homolog in the worm Caenorhabditis elegans, called C. elegans HCF (CeHCF). Loss of HCF- 1 function in a mammalian cell line and CeHCF function in worm leads to similar defects in cytokinesis and mitotic histone phosphorylation - defects that suggest disruption o the Aurora B kinase-Protein Phosphatase 1 (PP1) pathway. At least some of the defects caused by loss o HCF-1 function can be overcome by inactivation of the pRb tumor suppressor, suggesting that a natural role o HCF-1 is to suppress pRb function, either directly or indirectly. Thus, HCF-1 appears to be a key regulator o cell proliferation. In this project, we will continue to use innovative and complementary genetic, molecula and cell biological, and biochemical approaches to dissect the structure and function of HCF proteins in thei critical roles in cell division and proliferation. Our specific aims are (i) to study the roles of HCF-1 i mammalian-cell proliferation and differentiation; (ii) to identify and characterize the effectors of HCFfunction in mammalian cells; and to elucidate the roles of HCF-protein function in Caenorhabditis elegans.

An award has been made to California State University-Long Beach under the direction of Dr. Bruno Pernet for the acquisition of a laser scanning confocal microscope for research and teaching. The new microscope will support studies of cells and their internal structures at a finer resolution than currently possible. The microscope will support a dozen faculty members in biology, chemistry, and psychology, including several newly hired faculty. Research that will benefit includes studies of growth and development of fungal and mammalian cells, development of grass plants, gene expression in muscle cells, and neural structure and function in birds. The institution has a large Hispanic population, who can be expected to participate in research projects and to benefit from the instrument in their classes. Graduate students will use the instrument in their Masters Degree research.