Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2.
Vps4-A (vacuolar protein sorting 4-A) is a binding partner for a novel Rho family GTPase, Rnd2.
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Vps4-A（液泡蛋白分选 4-A）是新型 Rho 家族 GTPase Rnd2 的结合伴侣。
DOI：
10.1042/bj20020062
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发表时间：
2002
期刊：
影响因子：
--
通讯作者：
M. Negishi
中科院分区：
文献类型：
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作者：
Hiroko Tanaka;H. Fujita;H. Katoh;K. Mori;M. Negishi
Rho家族GTP酶参与多种生物学活动，包括胞内囊泡运输。RND2是Rho家族GTP酶的新成员，但其生物学功能尚不清楚。在本研究中，我们以RND2为诱饵进行了酵母双杂交筛选，发现RND2特异性地与Vps4-A(其中Vsp4-A是空泡蛋白分类4-A)结合，Vps4-A是AAA ATPase家族的成员，也是早期内体运输的中央调节因子。这种相互作用是通过酵母双杂交系统、体外结合和免疫共沉淀研究确定的。Vps4-A与鸟苷5‘-[β-硫代]三磷酸结合的活性形式和鸟苷5’-[β-硫代]二磷酸结合的非活性形式的RND2相关联。在HeLa细胞中表达的ATPase缺陷的Vps4-A突变体Vps4-A(E228Q)在早期内吞体内积聚。当RND2与Vps4-A(E228Q)共表达时，RND2被募集到Vps4-A结合的早期内吞体内。这些结果表明，RND2通过与Vps4-A直接结合参与调节内体转运。
Rho family GTPases are implicated in a variety of biological activities, including endocytic vesicle trafficking. Rnd2 is a new member of Rho family GTPases, but its biological functions are not known. In the present study, we have performed a yeast two-hybrid screening using Rnd2 as bait and revealed that Rnd2 binds specifically to Vps4-A (where Vsp4-A is vacuolar protein sorting 4-A), a member of the AAA ATPase family and a central regulator for early endosome trafficking. This interaction was determined by the yeast two-hybrid system, in vitro binding and co-immunoprecipitation studies. Vps4-A associated with both guanosine 5'-[beta-thio]triphosphate-bound active and guanosine 5'-[beta-thio]diphosphate-bound inactive forms of Rnd2. An ATPase-defective Vps4-A mutant, Vps4-A(E228Q), expressed in HeLa cells was accumulated in the early endosomes. When Rnd2 was co-expressed with Vps4-A(E228Q), Rnd2 was recruited to the Vps4-A-bound early endosomes. These results suggest that Rnd2 is involved in the regulation of endosomal trafficking via direct binding to Vps4-A.