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MBC in Press, published online ahead of print May 7, 2008
Mol. Biol. Cell 10.1091/mbc.E07-10-1069

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Submitted on October 24, 2007
Revised on April 1, 2008
Accepted on April 29, 2008

A Multi-Component Assembly Pathway Contributes to the Formation of Acentrosomal Microtubule Arrays in Interphase Drosophila cells

Gregory C. Rogers,* Nasser M. Rusan,* Mark Peifer,{dagger} and Stephen L. Rogers{ddagger}

*Department of Biology, {dagger}Lineberger Comprehensive Cancer Center, and {ddagger}Carolina Center for Genome Sciences, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599

Monitoring Editor: Tim Stearns

In animal cells, centrosomes nucleate microtubules that form polarized arrays to organize the cytoplasm. Drosophila presents an interesting paradox however, as centrosome-deficient mutant animals develop into viable adults. To understand this discrepancy, we analyzed behaviors of centrosomes and microtubules in Drosophila cells, in culture and in vivo, using a combination of live-cell imaging, electron microscopy, and RNAi. The canonical model of the cycle of centrosome function in animal cells states that centrosomes act as microtubule-organizing centers throughout the cell cycle. Unexpectedly, we found that many Drosophila cell-types display an altered cycle, in which functional centrosomes are only present during cell division. On mitotic exit, centrosomes disassemble producing interphase cells containing centrioles that lack microtubule-nucleating activity. Furthermore, steady-state interphase microtubule levels are not changed by codepleting both {gamma}-tubulins. However, {gamma}-tubulin RNAi delays microtubule regrowth after depolymerization, suggesting that it may function partially redundantly with another pathway. Therefore, we examined additional microtubule nucleating factors and found that Mini-spindles, CLIP-190, EB1, or dynein RNAi also delayed microtubule regrowth; surprisingly, this was not further prolonged when we codepleted {gamma}-tubulins. Taken together, these results modify our view of the cycle of centrosome function and reveal a multi-component acentrosomal microtubule assembly pathway to establish interphase microtubule arrays in Drosophila.

Address correspondence to: Stephen L. Rogers (srogers{at}bio.unc.edu)






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