title: ITSN2L Interacts with and Negatively Regulates RABEP1.
abstract: Intersectin-2Long (ITSN2L) is a multi-domain protein participating in endocytosis and exocytosis. In this study, RABEP1 was identified as a novel ITSN2L interacting protein using a yeast two-hybrid screen from a human brain cDNA library and this interaction, specifically involving the ITSN2L CC domain and RABEP1 CC3 regions, was further confirmed by in vitro GST (glutathione-S-transferase) pull-down and in vivo co-immunoprecipitation assays. Corroboratively, we observed that these two proteins co-localize in the cytoplasm of mammalian cells. Furthermore, over-expression of ITSN2L promotes RABEP1 degradation and represses RABEP1-enhanced endosome aggregation, indicating that ITSN2L acts as a negative regulator of RABEP1. Finally, we showed that ITSN2L and RABEP1 play opposite roles in regulating endocytosis. Taken together, our results indicate that ITSN2L interacts with RABEP1 and stimulates its degradation in regulation of endocytosis.
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. yangxiaoxu@hunnu.edu.cn.
authors.forename: Xiaoxu
authors.initials: X
authors.lastname: Yang
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. yanfeng1944@aliyun.com.
authors.forename: Feng
authors.initials: F
authors.lastname: Yan
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. huzech@gmail.com.
authors.forename: Zhicheng
authors.initials: Z
authors.lastname: He
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. ls198804180303@sina.com.
authors.forename: Shan
authors.initials: S
authors.lastname: Liu
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. chengfily@gmail.com.
authors.forename: Yeqing
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. weike8629@163.com.
authors.forename: Ke
authors.initials: K
authors.lastname: Wei
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. qingfusanduan@163.com.
authors.forename: Shiquan
authors.initials: S
authors.lastname: Gan
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. yuanjing27@126.com.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. wangshang887@gmail.com.
authors.forename: Shang
authors.initials: S
authors.lastname: Wang
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. xy5488@163.com.
authors.forename: Ye
authors.initials: Y
authors.lastname: Xiao
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. kaiqunren@126.com.
authors.forename: Kaiqun
authors.initials: K
authors.lastname: Ren
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. liuning0731@126.com.
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. huxiang@hunnu.edu.cn.
authors.forename: Xiang
authors.initials: X
authors.lastname: Hu
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. fengxiaoding@hotmail.com.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Ding
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. lebithu@foxmail.com.|Department of Dermatology, Xiangya Hospital, Central South University, Changsha 410081, China. lebithu@foxmail.com.
authors.forename: Xingwang
authors.initials: X
authors.lastname: Hu
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of State Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, China. xshlin@hunnu.edu.cn.
authors.forename: Shuanglin
authors.initials: S
authors.lastname: Xiang
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D047468:Immunoprecipitation; D042541:Intracellular Space / Q000378:metabolism; D019151:Peptide Library; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D025941:Protein Interaction Mapping; D021381:Protein Transport; D059748:Proteolysis; D020798:Two-Hybrid System Techniques; D033921:Vesicular Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C509776:ITSN2 protein, human; D019151:Peptide Library; C097227:RABEP1 protein, human; D033921:Vesicular Transport Proteins
keywords: ITSN2L; RABEP1; endocytosis; endosome; interactions