Tanji Takahiro, Nishikori Kenji, Haga Syoko, Kanno Yuki, Kobayashi Yusuke, Takaya Mai, Gengyo-Ando Keiko, Mitani Shohei, Shiraishi Hirohisa, Ohashi-Kobayashi Ayako
Department of Immunobiology, School of Pharmacy, Iwate Medical University, 2-1-1 Nishi-tokuta, Yahaba, Shiwa-gun, Iwate, 028-3694, Japan.
Department of Physiology, School of Medicine, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan.
BMC Cell Biol. 2016 Jan 27;17:4. doi: 10.1186/s12860-015-0076-2.
The intestinal cells of Caenorhabditis elegans are filled with heterogeneous granular organelles that are associated with specific organ functions. The best studied of these organelles are lipid droplets and acidified gut granules associated with GLO-1, a homolog of the small GTPase Rab38. In this study, we characterized a subset of the intestinal granules in which HAF-4 and HAF-9 localize on the membrane. HAF-4 and HAF-9 are ATP-binding cassette (ABC) transporter proteins that are homologous to the mammalian lysosomal peptide transporter TAPL (transporter associated with antigen processing-like, ABCB9).
Using transgenic worms expressing fluorescent protein-tagged marker proteins, we demonstrated that the HAF-4- and HAF-9-localizing organelles are not lipid droplets and do not participate in yolk protein transport. They were also ruled out as GLO-1-positive acidified gut granules. Furthermore, we clarified that the late endosomal protein RAB-7 localizes to the HAF-4- and HAF-9-localizing organelles and is required for their biogenesis.
Our results indicate that the HAF-4- and HAF-9-localizing organelles are distinct intestinal organelles associated with the endocytic pathway.
秀丽隐杆线虫的肠道细胞充满了与特定器官功能相关的异质性颗粒细胞器。这些细胞器中研究得最透彻的是脂滴和与小GTP酶Rab38的同源物GLO-1相关的酸化肠道颗粒。在本研究中,我们对HAF-4和HAF-9定位于膜上的一部分肠道颗粒进行了表征。HAF-4和HAF-9是ATP结合盒(ABC)转运蛋白,与哺乳动物溶酶体肽转运蛋白TAPL(与抗原加工样相关的转运蛋白,ABCB9)同源。
使用表达荧光蛋白标记的标记蛋白的转基因蠕虫,我们证明了HAF-4和HAF-9定位的细胞器不是脂滴,不参与卵黄蛋白运输。它们也被排除为GLO-1阳性的酸化肠道颗粒。此外,我们阐明了晚期内体蛋白RAB-7定位于HAF-4和HAF-9定位的细胞器,并且是其生物发生所必需的。
我们 的结果表明,HAF-4和HAF-9定位的细胞器是与内吞途径相关的独特肠道细胞器。