Guo Feng, Yoon Gyeong Mee, McCubbin Andrew G
Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.
Department of Biology University of North Carolina, Chapel Hill, NC 27599, USA.
Plants (Basel). 2013 Mar 4;2(1):72-86. doi: 10.3390/plants2010072.
Petunia inflata small CDPK-interacting protein 1 (PiSCP1) was identified as a pollen expressed PiCDPK1 interacting protein using the yeast two hybrid system and the interaction confirmed using pull-down and phosphorylation assays. PiSCP1 is pollen specific and shares amino acid homology with uncharacterized proteins from diverse species of higher plants, but no protein of known function. Expression of PiSCP1-GFP in vivo inhibited pollen tube growth and was shown to localize to peroxisomes in growing pollen tubes. As PiCDPK1 is plasma membrane localized, we investigated the localization of a second isoform, PiCDPK2, and show that it co-localizes to peroxisomes with PiSCP1 and that the two proteins interact in the yeast 2 hybrid interaction assay, suggesting that interaction with the latter CDPK isoform is likely the one of biological relevance. Both PiCDPK2 and PiSCP1 affect pollen tube growth, presumably by mediating peroxisome function, however how they do so is currently not clear.
利用酵母双杂交系统,在矮牵牛中鉴定出一种小的钙依赖蛋白激酶相互作用蛋白1(PiSCP1),它是一种在花粉中表达的与PiCDPK1相互作用的蛋白,并通过下拉和磷酸化试验证实了这种相互作用。PiSCP1是花粉特异性的,与多种高等植物中未鉴定的蛋白具有氨基酸同源性,但没有已知功能的蛋白。体内PiSCP1 - GFP的表达抑制花粉管生长,并显示定位于生长中的花粉管的过氧化物酶体。由于PiCDPK1定位于质膜,我们研究了第二种异构体PiCDPK2的定位,结果表明它与PiSCP1共定位于过氧化物酶体,并且这两种蛋白在酵母双杂交相互作用试验中相互作用,这表明与后一种钙依赖蛋白激酶异构体的相互作用可能具有生物学相关性。PiCDPK2和PiSCP1都影响花粉管生长,推测是通过介导过氧化物酶体功能,但目前它们如何发挥作用尚不清楚。