Comprehensive interference RNAi-sensitive Drosophila cells provide insights into lamella formation
and motor protein specificity By David Secko September 18, 2003
RNA interference (RNAi) is a molecular biological technique that has been extensively used to
deplete specific components in cells and has been invaluable in determining the requirements of a
given protein in a particular cellular process. One such system is the Drosophila S2 cell system,
which is very sensitive to double-stranded RNA (dsRNA)-mediated gene silencing, but comprehensive
study of the molecular components required for cellular processes in higher eukaryotes-such as
mitosis and cell migration-has been more difficult to examine. Two papers from the University of
California, San Francisco in the September 15 Journal of Cell Biology report comprehensive studies
elucidating the molecular requirements for actin-binding proteins in lamella formation and motor
proteins in mitosis.
In the first paper, Steven L. Rogers and colleagues used RNAi to investigate the requirements of
approximately 90 actin-binding proteins in lamella formation in the Drosophila S2 cell line. S2
cells are normally spherical, nonmotile, and have no polarity, but when plated on concanavalin A (con-A)-
covered glass coverslips, they undergo substantial morphological changes. The authors plated GFP-actin-
expressing S2 cells on con-A coverslips, examined them by fluorescence microscopy, and observed that
90% of the cells formed lamella. This process of lamella formation was used as an assay to test the
actin-binding proteins involved, and 19 of the proteins tested were shown to result in aberrant
lamella formations when depleted from cells. These included Arp2/3, SCAR, cofilin, Aip1, profilin,
and cyclase-associated protein (Journal of Cell Biology, 162:1079-1088, September 15, 2003).
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