Faculty Advisor or Committee Member

Mitsuo Ikebe, Advisor

Faculty Advisor or Committee Member

Joseph C. Bagshaw, Committee Member

Faculty Advisor or Committee Member

Daniel G. Gibson III, Committee Member

Faculty Advisor or Committee Member

Thomas W. Honeyman, Committee Member

Faculty Advisor or Committee Member

George Witman, Committee Member

Identifier

etd-061605-150140

Abstract

"The class IX myosin is a member of the myosin superfamily and found in variety of tissues. Myosin IX is quite unique among the myosin superfamily in that the tail region contains a GTPase activating protein (GAP) domain for the small GTP-binding protein, Rho. Recently it was reported that myosin IX shows processive movement that travels on an actin filament for a long distance. This was an intriguing discovery, because myosin IX is a “single-headed” myosin unlike other processive myosins which have “double-headed” structure. It has been thought that “processive” motors walk on their track with their two heads, thus traveling for a long distance. Therefore, it is reasonable to expect that the processive movement of single headed myosin IX is based on the unique feature of myosin IX motor function. In this study, I investigated the mechanism of processive movement of single-headed myosins by analyzing the mechanism of ATPase cycle of myosin IX that is closely correlated with the cross-bridge cycle (the mechanical cycle of actomyosin). In the first part, I performed the transient enzyme kinetic analysis of myosin IX using the motor domain construct to avoid the complexity raised by the presence of the tail domain. It was revealed that the kinetical characteristics of myosin IX ATPase is quite different from other processive myosins. It was particularly notable that the affinity of the weak actin binding state of Myosin IX was extremely high comparing with known myosins. It is thought that the high affinity for actin throughout the ATPase cycle is a major component to explain the processive movement of myosin IX. In the second part of this study, I cloned full length human myosin IX construct to further investigate the regulation of motor activity of myosin IX. It was revealed that the basal ATPase activity but not the actin dependent ATPase activity of myosin IX is inhibited by its tail region. Furthermore full-length myosin IX is regulated by calcium, presumably due to the calcium binding to the CaM light chain. These result suggest that the tail domain serves as a regulatory component of myosin IX."

Publisher

Worcester Polytechnic Institute

Degree Name

PhD

Department

Biology & Biotechnology

Project Type

Dissertation

Date Accepted

2005-06-16

Accessibility

Unrestricted

Subjects

motor protein, myosin superfamily

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