Cloning and characterization of betaCAP73, a novel regulator of beta-actin assembly.
In non-muscle cells, the isoactins are differentially localized, with beta-actin specifically enriched at the cell cortex within motile structures, such as lamellae, while gamma-actin shows no specific localization. To understand the sorting and regulation of beta-actin within moving cells, we previously isolated betaCAP73, a novel beta-actin-specific binding protein (Cell Motil. Cytoskel. 35 (1996) 175). Here, we have cloned and characterized the 4718 nucleotide betaCAP73 cDNA from an endothelial cell library. betaCAP73 cDNA contains six predicted ankyrin-like repeats at the amino terminus and is partially homologous to three previously reported sequences of unknown function. Northern analysis reveals betaCAP73 expression in all tissues tested, with highest levels in skeletal muscle. Consistent with previously demonstrated interactions between native betaCAP73 and beta-actin filament barbed-ends, recombinant betaCAP73 inhibits pyrene-actin assembly in an isoactin-specific manner. Compared to stationary cells betaCAP73 mRNA is down regulated in crawling cells. Similarly, motility-defective cells have increased betaCAP73 protein. Overexpression of full-length betaCAP73 induces the formation of novel membrane protrusions that are enriched in betaCAP73, while overexpression of betaCAP73 domains alters cell morphology. Combined, these results indicate that betaCAP73 modulates isoactin dynamics to regulate the morphological alterations required for cell growth and motility.
Welch
Alice Y
AY
Department of Cellular and Molecular Physiology, Tufts University School of Medicine, Boston, MA 02111, USA. welcha@usa.redcross.org
Herman
Ira M
IM
D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D017089:Ankyrin Repeat; D001483:Base Sequence; D019556:COS Cells; D002417:Cattle; D002465:Cell Movement; D048429:Cell Size; D002478:Cells, Cultured; D002645:Chickens; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D008840:Microfilament Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D000199:Actins; D018076:DNA, Complementary; D008840:Microfilament Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins