HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 281, Issue 47, 36347-36359, November 24, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/47/36347    most recent M607595200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Canton, D. A. Articles by Litchfield, D. W. Search for Related Content PubMed PubMed Citation Articles by Canton, D. A. Articles by Litchfield, D. W. Social Bookmarking                      What's this?

The Role of CKIP-1 in Cell Morphology Depends on Its Interaction with Actin-capping Protein^*

David A. Canton¹, Mary Ellen K. Olsten², Hanspeter Niederstrasser, John A. Cooper, and David W. Litchfield³

From the Regulatory Biology and Functional Genomics Research Group, Siebens-Drake Medical Research Institute, Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario N6A 5C1, Canada, and the Department of Cell Biology and Physiology, Washington University in St. Louis, St. Louis, Missouri 63110

CKIP-1 is a pleckstrin homology domain-containing protein that induces alterations of the actin cytoskeleton and cell morphology when expressed in human osteosarcoma cells. CKIP-1 interacts with the heterodimeric actin-capping protein in cells, so we postulated that this interaction was responsible for the observed cytoskeletal and morphological effects of CKIP-1. To test this postulate, we used peptide "walking arrays" and alignments of CKIP-1 with CARMIL, another CP-binding protein, to identify Arg-155 and Arg-157 of CKIP-1 as residues potentially required for its interactions with CP. CKIP-1 mutants harboring Arg-155 and Arg-157 substitutions exhibited greatly decreased CP binding, while retaining wild-type localization, the ability to interact with protein kinase CK2, and self-association. To examine the phenotype associated with expression of these mutants, we generated tetracycline-inducible human osteosarcoma cells lines expressing R155E,R157E mutants of CKIP-1. Examination of these cell lines reveals that CKIP-1 R155E,R157E did not induce the distinct changes in cell morphology and the actin cytoskeleton that are characteristic of wild-type CKIP-1 demonstrating that the interaction between CKIP-1 and CP is required for these cellular effects.

Received for publication, August 9, 2006 , and in revised form, August 30, 2006.

^* This work was supported in part by operating grants from the Canadian Institutes of Health Research (CIHR, Grant MOP 37854 to D. W. L.) and by National Institutes of Health Grant GM38542 (to J. A. C.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement"in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ Supported by a Canadian Graduate Scholarship from the CIHR.

² Supported by a CIHR Doctoral Award.

³ To whom correspondence should be addressed: Dept. of Biochemistry, University of Western Ontario, Medical Sciences Bldg., London, Ontario N6A 5C1, Canada. Tel.: 519-661-4186; Fax: 519-661-3175; E-mail: litchfi{at}uwo.ca.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				P. J. Vanderzalm, A. Pandey, M. E. Hurwitz, L. Bloom, H. R. Horvitz, and G. Garriga C. elegans CARMIL negatively regulates UNC-73/Trio function during neuronal development Development, April 1, 2009; 136(7): 1201 - 1210. [Abstract] [Full Text] [PDF]

				K. Kim, M. E. McCully, N. Bhattacharya, B. Butler, D. Sept, and J. A. Cooper Structure/Function Analysis of the Interaction of Phosphatidylinositol 4,5-Bisphosphate with Actin-capping Protein: IMPLICATIONS FOR HOW CAPPING PROTEIN BINDS THE ACTIN FILAMENT J. Biol. Chem., February 23, 2007; 282(8): 5871 - 5879. [Abstract] [Full Text] [PDF]