Cell
Volume 70, Issue 1, 10 July 1992, Pages 81-92
Journal home page for Cell

An actin-binding site containing a conserved motif of charged amino acid residues is essential for the morphogenic effect of villin

https://doi.org/10.1016/0092-8674(92)90535-KGet rights and content

Abstract

The actin-binding protein villin induces microvillus growth and reorganization of the cytoskeleton in cells that do not normally produce this protein. Transfection of mutagenized villin cDNAs Into CV-1 cells was used to show that a conserved, COOH-terminally located cluster of charged amino acid residues (KKEK) is crucial for the morphogenic activity of villin in vivo. In vitro experiments with a 22 amino acid synthetic peptide corresponding to this region of villin provide evidence that this motif is part of an F-actin-binding site that induces G-actin to polymerize. Chemical cross-linking of actin to this peptide, the effects of amino acid substitutions in peptides, and the behavior of villin variants further corroborate the participation of the KKEK sequence in actin contacts.

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      Citation Excerpt :

      Oligomers also do not bind this domain because CaM and actin had additive effects on PMCA catalytic activity (Fig. 8). A possible candidate for the binding site may be the positively charged acidic phospholipid site in the PMCA, because actin is negatively charged at physiological pH. Lysine-rich clusters in actin-binding proteins have been identified to be involved in actin binding, e.g. the DAIKKK sequence in actin depolymerization factor, cofilin, and tropomyosin (89); the KKGGKKKG sequence of myosin (90); the KSKLKKT sequence in thymosin β4 (91), and the sequence KKEK in villin (92). Although there is no conserved sequence for actin binding between the different actin-binding proteins, a consistent motif rich in basic residues exists, which is also present in the PMCA sequence at the cytosolic loop between the transmembrane segments 2 and 3.

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