title: RC/BTB2 is essential for formation of primary cilia in mammalian cells.
abstract: RC/BTB2 is a binding partner of sperm associated antigen 16S (SPAG16S), which is a regulator of spermiogenesis in mice, a process during which sperm flagella are formed. The expression of Rc/btb2 is also regulated by multicilin, a protein that controls ciliogenesis. Given that mouse Rc/btb2 mRNA is not only expressed in tissues bearing motile cilia, but also in tissues without motile cilia, we investigated whether RC/BTB2 plays a role in the general process of ciliogenesis by studying two cell lines that have primary cilia, NIH3T3, and IMCD3. We discovered that the subcellular localization of RC/BTB2 in the NIH3T3 and IMCD3 cells encompasses the pathway for ciliogenesis. RC/BTB2 was found in the Golgi bodies and centrosomes, two key structures essential for normal ciliogenesis. Knockdown of Rc/btb2 gene expression in these cell lines disrupted ciliogenesis. The percentage of cells with primary cilia was significantly reduced in stable cell lines transduced with specific Rc/btb2 shRNA viruses as compared to the control cells. When cilia were formed in the knockdown cells, they were significantly shorter than those in the control cells. Knockdown of Rc/btb2 expression did not affect cell proliferation and the cell cycle. Exogenous expression of RC/BTB2 in these stable knockdown cells restored ciliogenesis. These findings suggest that RC/BTB2 is a necessary component of the process of formation of primary cilia in somatic cells, perhaps through the transportation of cargos from Golgi bodies to centrosomes for cilia assembling.
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei, China.|Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Ling
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Radiation Medicine, Second Military Medical University, Shanghai, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Ni
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Jinghua
authors.initials: J
authors.lastname: Wu
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei, China.|Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Junping
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Infectious Diseases, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
authors.forename: Zhengang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.|Department of Dermatology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Hongfei
authors.initials: H
authors.lastname: Li
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei, China.
authors.forename: Yuqin
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Maria E
authors.initials: ME
authors.lastname: Teves
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei, China.
authors.forename: Shizheng
authors.initials: S
authors.lastname: Song
authors.affiliation: Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Jerome F
authors.initials: JF
authors.lastname: Strauss
authors.affiliation: School of Public Health, Wuhan University of Science and Technology, Wuhan, Hubei, China.|Department of Obstetrics & Gynecology, Virginia Commonwealth University, Richmond, Virginia.
authors.forename: Zhibing
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D000818:Animals; D018385:Centrosome / Q000378:metabolism*; D002923:Cilia / Q000235:genetics / Q000378:metabolism; D003598:Cytoskeletal Proteins / Q000096:biosynthesis* / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D051379:Mice; D041681:NIH 3T3 Cells
chemical_list: D003598:Cytoskeletal Proteins
keywords: Golgi body; RC/BTB2; centriole; ciliogenesis