© The Rockefeller University Press,
0021-9525/1998//121 $5.00
The Journal of Cell Biology, Volume 143, Number 1,
, 1998 121-133
Why Are Two Different Cross-linkers Necessary for Actin Bundle Formation In Vivo and What Does Each Cross-link Contribute?
Lewis G. Tilney,
Patricia S. Connelly,
Kelly A. Vranich,
Michael K. Shaw, and
Gregory M. Guild
Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
In developing Drosophila bristles two species of cross-linker, the forked proteins and fascin, connect adjacent actin filaments into bundles. Bundles form in three phases: (a) tiny bundles appear; (b) these bundles aggregate into larger bundles; and (c) the filaments become maximally cross-linked by fascin. In mutants that completely lack forked, aggregation of the bundles does not occur so that the mature bundles consist of <50 filaments versus
700 for wild type. If the forked concentration is genetically reduced to half the wild type, aggregation of the tiny bundles occurs but the filaments are poorly ordered albeit with small patches of fascin cross-linked filaments. In mutants containing an excess of forked, all the bundles tend to aggregate and the filaments are maximally crossbridged by fascin. Alternatively, if fascin is absent, phases 1 and 2 occur normally but the resultant bundles are twisted and the filaments within them are poorly ordered. By extracting fully elongated bristles with potassium iodide which removes fascin but leaves forked, the bundles change from being straight to twisted and the filaments within them become poorly ordered. From these observations we conclude that (a) forked is used early in development to aggregate the tiny bundles into larger bundles; and (b) forked facilitates fascin entry into the bundles to maximally cross-link the actin filaments into straight, compact, rigid bundles. Thus, forked aligns the filaments and then directs fascin binding so that inappropriate cross-linking does not occur.
Key Words: actin Drosophila bristles cross-links fascin forked
Many thanks to N. Petersen and L. Cooley for their gifts of fly strains and antibodies against the forked protein and fascin, respectively. We are especially grateful to M. Mooseker who for two summers gave advice, encouragement, space, and equipment while L.G. Tilney was in Woods Hole. We would also like to thank the reviewers of this manuscript for fair and just criticism, and our editor D. DeRosier (Brandeis University) for helping to improve the manuscript.
Address all correspondence to G. Guild, Department of Biology, University of Pennsylvania, Philadelphia, Pennsylvania 19104. Tel.: (215) 898-6388. Fax: (215) 898-8780. E-mail: gguild{at}sas.upenn.edu
1. Abbreviation used in this paper: KI, potassium iodide.

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