Overexpression of human fibroblast caldesmon fragment containing actin- , Ca++/calmodulin-, and tropomyosin-binding domains stabilizes endogenous tropomyosin and microfilaments

doi:10.1083/jcb.125.2.359

This Article

	Full Text (PDF, 3035K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Warren, K. S.
	Articles by Lin, J. J.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Warren, K. S.
	Articles by Lin, J. J.


Social Bookmarking

	What's this?

ARTICLES

Overexpression of human fibroblast caldesmon fragment containing actin- , Ca++/calmodulin-, and tropomyosin-binding domains stabilizes endogenous tropomyosin and microfilaments

KS Warren, JL Lin, DD Wamboldt and JJ Lin
Department of Biological Sciences, University of Iowa, Iowa City 52242- 1324.

Fibroblast caldesmon is a protein postulated to participate in the modulation of the actin cytoskeleton and the regulation of actin-based motility. The cDNAs encoding the NH2-terminal (aa.1-243, CaD40) and COOH-terminal (aa.244-538, CaD39) fragments of human caldesmon were subcloned into expression vectors and we previously reported that bacterially produced CaD39 protein retains its actin-binding properties as well as its ability to enhance low M(r) tropomyosin (TM) binding to actin and to inhibit TM-actin-activated HMM ATPase activity in vitro (Novy, R. E., J. R. Sellers, L.-F. Liu, and J. J.-C. Lin. 1993. Cell Motil. Cytoskeleton. 26:248-261). Bacterially produced CaD40 does not bind actin. To study the in vivo effects of CaD39 expression on the stability of actin filaments in CHO cells, we isolated and characterized stable CHO transfectants which express varying amounts of CaD39. We found that expression of CaD39 in CHO cells stabilized microfilament bundles as well as endogenous TM. CaD39-expressing clones displayed an increased resistance to cytochalasin B and Triton X-100 treatments and yielded increased amounts of TM-containing actin filaments in microfilament isolation procedures. In addition, analysis of these clones with immunoblotting and indirect immunofluorescence microscopy with anti-TM antibody revealed that stabilized endogenous TM and enhanced TM-containing microfilament bundles parallel increased amounts of CaD39 expression. The increased TM observed corresponded to a decrease in TM turnover rate and did not appear to be due to increased synthesis of endogenous TM. Additionally, the phenomenon of stabilized TM did not occur in stable CHO clones expressing CaD40. Therefore, it is likely that CaD39 can enhance TM's binding to F-actin in vivo, thus reducing TM's rate of turnover and stabilizing actin microfilament bundles.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Mayanagi, T., Morita, T., Hayashi, K., Fukumoto, K., Sobue, K. (2008). Glucocorticoid Receptor-mediated Expression of Caldesmon Regulates Cell Migration via the Reorganization of the Actin Cytoskeleton. J. Biol. Chem. 283: 31183-31196 [Abstract] [Full Text]
Gunning, P., O'neill, G., Hardeman, E. (2008). Tropomyosin-Based Regulation of the Actin Cytoskeleton in Time and Space. Physiol. Rev. 88: 1-35 [Abstract] [Full Text]
Hossain, M. M., Crish, J. F., Eckert, R. L., Lin, J. J.-C., Jin, J.-P. (2005). h2-calponin Is Regulated by Mechanical Tension and Modifies the Function of Actin Cytoskeleton. J. Biol. Chem. 280: 42442-42453 [Abstract] [Full Text]
Cuomo, M. E., Knebel, A., Platt, G., Morrice, N., Cohen, P., Mittnacht, S. (2005). Regulation of Microfilament Organization by Kaposi Sarcoma-associated Herpes Virus-cyclin{middle dot}CDK6 Phosphorylation of Caldesmon. J. Biol. Chem. 280: 35844-35858 [Abstract] [Full Text]
Li, Y., Lin, J. L. C., Reiter, R. S., Daniels, K., Soll, D. R., Lin, J. J. C. (2004). Caldesmon mutant defective in Ca2+-calmodulin binding interferes with assembly of stress fibers and affects cell morphology, growth and motility. J. Cell Sci. 117: 3593-3604 [Abstract] [Full Text]
Yamashiro, S., Chern, H., Yamakita, Y., Matsumura, F. (2001). Mutant Caldesmon Lacking cdc2 Phosphorylation Sites Delays M-Phase Entry and Inhibits Cytokinesis. Mol. Biol. Cell 12: 239-250 [Abstract] [Full Text]
Wang, Z., Danielsen, A. J., Maihle, N. J., McManus, M. J. (1999). Tyrosine Phosphorylation of Caldesmon Is Required for Binding to the Shc{middle dot}Grb2 Complex. J. Biol. Chem. 274: 33807-33813 [Abstract] [Full Text]
Ishikawa, R., Yamashiro, S., Kohama, K., Matsumura, F. (1998). Regulation of Actin Binding and Actin Bundling Activities of Fascin by Caldesmon Coupled with Tropomyosin. J. Biol. Chem. 273: 26991-26997 [Abstract] [Full Text]
Kashiwada, K., Nishida, W., Hayashi, K.'i., Ozawa, K., Yamanaka, Y., Saga, H., Yamashita, T., Tohyama, M., Shimada, S., Sato, K., Sobue, K. (1997). Coordinate Expression of alpha -Tropomyosin and Caldesmon Isoforms in Association with Phenotypic Modulation of Smooth Muscle Cells. J. Biol. Chem. 272: 15396-15404 [Abstract] [Full Text]
Pelham, R., Lin, J., Wang, Y. (1996). A high molecular mass non-muscle tropomyosin isoform stimulates retrograde organelle transport. J. Cell Sci. 109: 981-989 [Abstract]
Pittenger, M., Kistler, A, Helfman, D. (1995). Alternatively spliced exons of the beta tropomyosin gene exhibit different affinities for F-actin and effects with nonmuscle caldesmon. J. Cell Sci. 108: 3253-3265 [Abstract]