POLYCYSTIN-1 AND POLYCYSTIN-2 FUNCTION SYNERGISTICALLY IN FISSION YEAST

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POLYCYSTIN-1 AND POLYCYSTIN-2 FUNCTION SYNERGISTICALLY IN FISSION YEAST

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Co-author 1

Takayuki Koyano koyano@shigei.or.jp Shigei Medical Research Institute Division of Cell Biology Okayama Japan *

Co-author 2

Kaori Onishi k-onishi@shigei.or.jp Shigei Medical Research Institute Division of Cell Biology Okayama Japan -

Co-author 3

Makoto Matsuyama matsuyama@shigei.or.jp Shigei Medical Research Institute Division of Molecular Genetics Okayama Japan -

Co-author 4

Koji Manabe manabe@shigei.or.jp Shigei Medical Research Hospital Shigei Medical Research Hospital Okayama Japan -

Co-author 5

Masaki Fukushima fukushima@shigei.or.jp Shigei Medical Research Institute Division of Molecular Genetics Okayama Japan -

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Introduction

Mutations in polycystins cause to autosomal dominant polycystic kidney disease (ADPKD), the most common genetic inheritant kidney disease. Although the functions of polycystins are critical to ADPKD progression, the details are still under debate. Here, we analyzed the fundamental functions of polycystins using a novel model system based on fission yeast. Fission yeast has a putative polycystin protein, Pkd2, whose gene deletion led to lethality.

Methods

We expressed the human genes encoding polycystin-2 (hPkd2) and polycystin-1 (hPkd1) ectopically in fission yeast. The CDRE (Calcineurin Dependent Response Element) reporter was used to evaluate cellular function.

Results

Although pkd2 overexpression activates the calcium signaling, the effect of hpkd2 overexpression is minor. Furthermore, hpkd2 do not complete the essentiality of pkd2 in fission yeast. Pkd2 localizes not only to the endoplasmic reticulum (ER), but also to the plasma membrane. In contrast, hPkd2 only localizes to the ER in fission yeast. These data suggest that simply expressing hPkd2 does not work well in fission yeast. Then, we sought to the conditions under which hPkd2 functions in fission yeast. To this end, we constructed several forms of hPkd2, including adding membrane targeting motif sequence. Finally, we found that expressing hPkd2 with hPkd1 effectively activates calcium signaling in fission yeast.

Conclusion

The function of hPkd2 is expressed in fission yeast through co-expression with hPkd1. We adapted this system to analyze other signaling pathways, localization, and pathogenic mutations. In this presentation, we will discuss the fundamental functions of human polycystins.

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