In vivo analysis of functional domains from villin and gelsolin.
Open Access
- 1 March 1992
- journal article
- research article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 116 (5), 1145-1155
- https://doi.org/10.1083/jcb.116.5.1145
Abstract
Transfected CV1 cells were used to compare the in vivo effects of various domains of villin and gelsolin. These two homologous actin modulating proteins both contain a duplicated severin-like sequence. Villin has in addition a carboxy-terminal domain, the headpiece, which accounts for its bundling activity. The effects of the villin-deleted mutants were compared with those of native villin. Our results show that essential domains of villin required to induce the growth of microvilli and F-actin redistribution are present in the first half of the core and in the headpiece. We also show that the second half of the villin core cannot be exchanged by its homolog in gelsolin. When expressed at high levels of CV1 cells, full length gelsolin completely disrupted stress fibers without change of the cell shape. Addition of the villin headpiece to gelsolin had no effect on the phenotype induced by gelsolin alone. Expression of the first half of gelsolin induced similar modifications as capping proteins and rapid cell mortality; this deleterious effect on the cell structure was also observed when the headpiece was linked to the first half of gelsolin. In cells expressing the second half of gelsolin, a dotted F-actin staining was often seen. Moreover elongated dorsal F-actin structures were observed when the headpiece was linked to the second gelsolin domain. These studies illustrate the patent in vivo severing activity of gelsolin as well as the distinct functional properties of villin core in contrast to gelsolin.Keywords
This publication has 42 references indexed in Scilit:
- Differential effects of gelsolins on tissue culture cellsCell Motility, 1990
- End-label fingerprintings show that the N- and C-termini of actin are in the contact site with gelsolinBiochemistry, 1989
- Gelsolin has three actin-binding sites.The Journal of cell biology, 1988
- Localization and mobility of gelsolin in cells.The Journal of cell biology, 1988
- Microinjection of gelsolin into living cells.The Journal of cell biology, 1987
- Tropomyosin distinguishes between the two actin-binding sites of villin and affects actin-binding properties of other brush border proteins.The Journal of cell biology, 1987
- Disruption of microfilament organization after injection of F-actin capping proteins into living tissue culture cellsNature, 1983
- Immunolocalization of the 110,000 molecular weight cytoskeletal protein of intestinal microvilliJournal of Molecular Biology, 1981
- “Western Blotting”: Electrophoretic transfer of proteins from sodium dodecyl sulfate-polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein AAnalytical Biochemistry, 1981
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970