Fujiki Yuki, Yoshimoto Kohki, Ohsumi Yoshinori
Department of Cell Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan.
Plant Physiol. 2007 Mar;143(3):1132-9. doi: 10.1104/pp.106.093864. Epub 2007 Jan 26.
Yeast (Saccharomyces cerevisiae) Atg6/Vps30 is required for autophagy and the sorting of vacuolar hydrolases, such as carboxypeptidase Y. In higher eukaryotes, however, roles for ATG6/VPS30 homologs in vesicle sorting have remained obscure. Here, we show that AtATG6, an Arabidopsis (Arabidopsis thaliana) homolog of yeast ATG6/VPS30, restored both autophagy and vacuolar sorting of carboxypeptidase Y in a yeast atg6/vps30 mutant. In Arabidopsis cells, green fluorescent protein-AtAtg6 protein localized to punctate structures and colocalized with AtAtg8, a marker protein of the preautophagosomal structure. Disruption of AtATG6 by T-DNA insertion resulted in male sterility that was confirmed by reciprocal crossing experiments. Microscopic analyses of AtATG6 heterozygous plants (AtATG6/atatg6) crossed with the quartet mutant revealed that AtATG6-deficient pollen developed normally, but did not germinate. Because other atatg mutants are fertile, AtAtg6 likely mediates pollen germination in a manner independent of autophagy. We propose that Arabidopsis Atg6/Vps30 functions not only in autophagy, but also plays a pivotal role in pollen germination.
酵母(酿酒酵母)中的Atg6/Vps30对于自噬以及液泡水解酶(如羧肽酶Y)的分选是必需的。然而,在高等真核生物中,ATG6/VPS30同源物在囊泡分选中的作用仍不清楚。在这里,我们表明AtATG6,即拟南芥中酵母Atg6/Vps30的同源物,在酵母atg6/vps30突变体中恢复了羧肽酶Y的自噬和液泡分选。在拟南芥细胞中,绿色荧光蛋白-AtAtg6蛋白定位于点状结构,并与自噬前体结构的标记蛋白AtAtg8共定位。通过T-DNA插入破坏AtATG6导致雄性不育,这一点通过正反交实验得到证实。对与四重突变体杂交的AtATG6杂合植物(AtATG6/atatg6)的显微镜分析表明,AtATG6缺陷型花粉发育正常,但不萌发。由于其他atatg突变体是可育的,AtAtg6可能以独立于自噬的方式介导花粉萌发。我们提出,拟南芥Atg6/Vps30不仅在自噬中起作用,而且在花粉萌发中起关键作用。