Overexpression of microfilament-stabilizing human caldesmon fragment, CaD39, affects cell attachment, spreading, and cytokinesis.
Previous studies have demonstrated that overexpression of the carboxyl-terminal fragment, CaD39, of human fibroblast caldesmon in Chinese hamster ovary cells protected endogenous tropomyosin from turnover and stabilized actin microfilament bundles [Warren et al., 1994: J. Cell Biol. 125:359-368]. To assess the consequences of having CaD39-stabilized microfilaments in living cell, we characterized the motile behaviors of stable CaD39-expressing lines. We here found that CaD39-expressing cells adhered faster to plastic, glass, fibronectin-coated glass, and collagen-coated glass than control cells. Moreover, the CaD39-expressing cells also exhibited enhanced spreading immediately after attachment. Despite these differences, overexpression of CaD39 had little effect on the velocity of intracellular granule movement, or the velocity and persistence of cellular translocation. However, CaD39-expressing cells were more elongate and encompassed less area than non-expressing cells during migration in a wound-healing assay. In interphase cells, the expressed CaD39 fragments were found associated with tropomyosin-enriched microfilaments. Like endogenous caldesmon, the CaD39 fragment was also modified at mitosis. Although a significant portion of CaD39 underwent only partial modification, the majority of the CaD39 was released from the microfilaments during mitosis. This is consistent with the finding that the CaD39-induced advantage for attachment and spreading was lost during mitosis. In CaD39-expressing cells, an incomplete release of the CaD39 from microfilaments at mitosis was found which may be responsible for the increase in the frequency of multinuclear cells in CaD39-expressing lines.
Warren
K S
KS
Department of Biological Sciences, University of Iowa, Iowa City 52242-1324, USA.
Shutt
D C
DC
McDermott
J P
JP
Lin
J L
JL
Soll
D R
DR
Lin
J J
JJ
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion; D002455:Cell Division; D002465:Cell Movement; D006224:Cricetinae; D015870:Gene Expression; D006801:Humans; D008938:Mitosis; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism
D002148:Calmodulin-Binding Proteins; D010446:Peptide Fragments