Yamada Koji, Hirotsu Takaaki, Matsuki Masahiro, Kunitomo Hirofumi, Iino Yuichi
Department of Biophysics and Biochemistry, Molecular Genetics Research Laboratory, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Genetics. 2009 Apr;181(4):1347-57. doi: 10.1534/genetics.108.099002. Epub 2009 Feb 2.
Caenorhabditis elegans genome carries two Ggamma genes, gpc-1 and gpc-2, and two Gbeta genes, gpb-1 and gpb-2. Of these, gpc-2 and gpb-1 are expressed ubiquitously and are essential for viability. Through a genetic screen, we identified gpc-1 as essential for olfactory adaptation. While wild-type animals show decreased chemotaxis to the odorant benzaldehyde after a short preexposure to the odorant, gpc-1 mutants are still attracted to the odorant after the same preexposure. Cell-specific rescue experiments show that gpc-1 acts in the AWC olfactory neurons. Coexpression of GPC-1 and GPB-1, but not GPB-2, caused enhanced adaptation, indicating that GPC-1 may act with GPB-1. On the other hand, knock down of gpc-2 by cell-targeted RNAi caused reduced chemotaxis to the odorant in unadapted animals, indicating that GPC-2 mainly act for olfactory sensation and the two Ggamma's have differential functions. Nonetheless, overexpression of gpc-2 in AWC neurons rescued the adaptation defects of gpc-1 mutants, suggesting partially overlapping functions of the two Ggamma's. We further tested genetic interaction of gpc-1 with several other genes involved in olfactory adaptation. Our analyses place goa-1 Goalpha and let-60 Ras in parallel to gpc-1. In contrast, a gain-of-function mutation in egl-30 Gqalpha was epistatic to gpc-1, suggesting the possibility that gpc-1 Ggamma may act upstream of egl-30 Gqalpha.
秀丽隐杆线虫基因组携带两个Gγ基因,即gpc-1和gpc-2,以及两个Gβ基因,即gpb-1和gpb-2。其中,gpc-2和gpb-1在全身表达,对生存能力至关重要。通过遗传筛选,我们确定gpc-1对嗅觉适应至关重要。野生型动物在短暂预接触气味剂苯甲醛后对该气味剂的趋化性降低,而gpc-1突变体在相同预接触后仍被该气味剂吸引。细胞特异性拯救实验表明,gpc-1在AWC嗅觉神经元中起作用。GPC-1和GPB-1(而非GPB-2)的共表达导致适应性增强,表明GPC-1可能与GPB-1共同起作用。另一方面,通过细胞靶向RNAi敲低gpc-2会导致未适应动物对气味剂的趋化性降低,表明GPC-2主要作用于嗅觉感知,且这两个Gγ具有不同的功能。尽管如此,在AWC神经元中过表达gpc-2挽救了gpc-1突变体的适应缺陷,表明这两个Gγ的功能部分重叠。我们进一步测试了gpc-1与其他几个参与嗅觉适应的基因的遗传相互作用。我们的分析表明,goa-1 Goα和let-60 Ras与gpc-1平行。相比之下,egl-30 Gqα的功能获得性突变对gpc-1是上位性的,这表明gpc-1 Gγ可能在egl-30 Gqα上游起作用。