Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella.
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.
Huang
Kaiyao
K
Department of Molecular Cell and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Diener
Dennis R
DR
Mitchell
Aaron
A
Pazour
Gregory J
GJ
Witman
George B
GB
Rosenbaum
Joel L
JL
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D002448:Cell Adhesion; D016825:Chlamydomonas reinhardtii / Q000378:metabolism*; D003001:Cloning, Molecular; D005407:Flagella / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D050396:TRPP Cation Channels / Q000737:chemistry* / Q000378:metabolism; D013696:Temperature
D049452:Green Fluorescent Proteins; D050396:TRPP Cation Channels