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MOLECULAR INSIGHTS INTO THE LOCALIZATION AND ROCK-BINDING ACTIVITIES OF THE CYTOSKELETAL PROTEIN SHROOM2

Zalewski, Jenna (2017) MOLECULAR INSIGHTS INTO THE LOCALIZATION AND ROCK-BINDING ACTIVITIES OF THE CYTOSKELETAL PROTEIN SHROOM2. Doctoral Dissertation, University of Pittsburgh. (Unpublished)

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Abstract

As an organism develops, it undergoes dramatic changes in shape as tissues and organs form. Changes in tissue morphology are driven at the cellular level by the coordinated movement, division, and shape change of constituent cells. These processes in turn are regulated by dynamic subcellular networks of actin and myosin. Rho Kinase (Rock) is a cytoskeletal regulator that phosphorylates non-muscle myosin II to drive remodeling of cellular architecture. Rock can be recruited to distinct subcellular locales via interaction with the Shroom (Shrm) family of proteins. Shrm proteins harbor several regulatory domains including an N-terminal PDZ that is used for localization, an SD1 that binds F-actin, and a C-terminal SD2 responsible for association with Rock. My thesis is focused on understanding the molecular mechanisms employed by these domains to contribute to Shrm function. To this end, I crystallized and determined the structure of the complex between SD2 and the Shrm binding domain (SBD) of Rock. This structure provides detailed insight into the interface between the two domains and motivated biochemical and in vivo probing to pinpoint residues that contribute to SD2-SBD recognition. Kinase activity assays indicate that Shrm proteins can stimulate Rock catalytic activity, demonstrating for the first time that Shrm proteins function directly as Rock activators. Additionally, I used structural and biochemical techniques to dissect the function of the Shrm2 PDZ domain and determined its binding partner and binding mechanism. These data enhance our understanding of how the Shrm protein family contributes to development.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Zalewski, Jennajkz3@pitt.edujkz30000-0002-9763-3028
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairVanDemark, Andrew P.andyv@pitt.eduANDYV
Committee MemberBerman, Andrea J.ajb190@pitt.eduAJB190
Committee MemberHildebrand, Jeffrey D.jeffh@pitt.eduJEFFH
Committee MemberLee, Tina H.thl@andrew.cmu.edu
Committee MemberPipas, James M.pipas@pitt.eduPIPAS
Date: 28 September 2017
Date Type: Publication
Defense Date: 14 July 2017
Approval Date: 28 September 2017
Submission Date: 7 August 2017
Access Restriction: 5 year -- Restrict access to University of Pittsburgh for a period of 5 years.
Number of Pages: 195
Institution: University of Pittsburgh
Schools and Programs: Dietrich School of Arts and Sciences > Biological Sciences
Degree: PhD - Doctor of Philosophy
Thesis Type: Doctoral Dissertation
Refereed: Yes
Uncontrolled Keywords: Shroom, Rho-Kinase, Structural Biology, coiled-coil, protein-protein interactions, PDZ, cytoskeleton
Date Deposited: 29 Sep 2017 01:19
Last Modified: 29 Sep 2017 01:19
URI: http://d-scholarship.pitt.edu/id/eprint/33068

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  • MOLECULAR INSIGHTS INTO THE LOCALIZATION AND ROCK-BINDING ACTIVITIES OF THE CYTOSKELETAL PROTEIN SHROOM2. (deposited 29 Sep 2017 01:19) [Currently Displayed]

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