NMR INVESTIGATION OF HUMAN CKSHS1

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. TITLE OF PROJECT: NMR Investigation of Human CksHs1 PILOT-PROJECT INVESTIGATOR: James J.A. Huntley, Ph.D. PERFORMANCE SITES: National Center for Genome Resources, The Scripps Research Institute, New Mexico State University The broad, long-term objective of this investigation is to determine the role of protein structural dynamics in the processes that constitute cell growth and division. The investigations specific aim is to explore and characterize the dynamics of a protein believed to have multiple roles, including one that serves to reinforce the correct timing and progression of the cell cycle. The protein, CksHs1, has been shown to bind to a number of proteins and inorganic molecules, including phosphate and Cyclin Dependent Kinase 2 (CDK2), a key enzyme involved in cell cycle regulation. Comparison of CksHs1 in various states has suggested that conformational flexibility of the protein is an important facet of its function. However, these investigations have not fully characterized this flexibility, or exactly how it relates to CksHs1 function. Without this information, a complete picture of CksHs1 function in the process of cell growth and division will not be possible. In order to better understand the role of dynamics in CksHs1 function, we will study the structure of CksHs1 by use of Nuclear Magnetic Resonance (NMR) relaxation techniques. Our central hypothesis is that CksHs1 displays novel features of dynamics that give the protein the ability to fold into slightly different structures depending upon the structure of the binding partner(s). We aim to characterize the dynamics of free CksHs1, the alteration to CksHs1 dynamics that occurs upon binding of phosphate, map CksHs1 and CDK2 interactions, and characterize the dynamics of the CksHs1-CDK2 binding process. The rationale for this research is that we will better understand the components and pathways of cell cycle control, and ultimately how features of protein structure dynamics may be exploited in the development of novel therapeutic agents or strategies to treat cancer or other diseases of uncontrolled cell growth.