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.
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5 November 2007
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November 05 2007
Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella
Kaiyao Huang,
Kaiyao Huang
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
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Dennis R. Diener,
Dennis R. Diener
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
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Aaron Mitchell,
Aaron Mitchell
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
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Gregory J. Pazour,
Gregory J. Pazour
2Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
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George B. Witman,
George B. Witman
3Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655
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Joel L. Rosenbaum
Joel L. Rosenbaum
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
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Kaiyao Huang
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
Dennis R. Diener
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
Aaron Mitchell
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
Gregory J. Pazour
2Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605
George B. Witman
3Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655
Joel L. Rosenbaum
1Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520
Correspondence to J.L. Rosenbaum: [email protected]
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.
Received:
April 13 2007
Accepted:
October 10 2007
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2007
J Cell Biol (2007) 179 (3): 501–514.
Article history
Received:
April 13 2007
Accepted:
October 10 2007
Citation
Kaiyao Huang, Dennis R. Diener, Aaron Mitchell, Gregory J. Pazour, George B. Witman, Joel L. Rosenbaum; Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella . J Cell Biol 5 November 2007; 179 (3): 501–514. doi: https://doi.org/10.1083/jcb.200704069
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