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Published online November 5, 2007
doi:10.1083/jcb.200704069
The Journal of Cell Biology, Vol. 179, No. 3, 501-514
The Rockefeller University Press, 0021-9525 $30.00
© 2007 Huang et al.
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Article

 Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella

Kaiyao Huang1, Dennis R. Diener1, Aaron Mitchell1, Gregory J. Pazour2, George B. Witman3, and Joel L. Rosenbaum1

1 Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
2 Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
3 Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655

Correspondence to J.L. Rosenbaum: joel.rosenbaum{at}yale.edu

To analyze the function of ciliary polycystic kidney disease 2 (PKD2) and its relationship to intraflagellar transport (IFT), we cloned the gene encoding Chlamydomonas reinhardtii PKD2 (CrPKD2), a protein with the characteristics of PKD2 family members. Three forms of this protein (210, 120, and 90 kD) were detected in whole cells; the two smaller forms are cleavage products of the 210-kD protein and were the predominant forms in flagella. In cells expressing CrPKD2–GFP, about 10% of flagellar CrPKD2–GFP was observed moving in the flagellar membrane. When IFT was blocked, fluorescence recovery after photobleaching of flagellar CrPKD2–GFP was attenuated and CrPKD2 accumulated in the flagella. Flagellar CrPKD2 increased fourfold during gametogenesis, and several CrPKD2 RNA interference strains showed defects in flagella-dependent mating. These results suggest that the CrPKD2 cation channel is involved in coupling flagellar adhesion at the beginning of mating to the increase in flagellar calcium required for subsequent steps in mating.

Abbreviations used in this paper: CrPKD2, Chlamydomonas reinhardtii PKD2; DIC, differential interference contrast; FMG, flagellar membrane glycoprotein; IFT, intraflagellar transport; mt, mating type; NaPPi, sodium pyrophosphate; PKD, polycystic kidney disease; TRP, transient receptor potential; UTR, untranslated region.


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