TY - JOUR
T1 - Novel localizations and interactions of intercellular bridge proteins revealed by proteomic profiling
AU - Iwamori, Tokuko
AU - Iwamori, Naoki
AU - Matsumoto, Masaki
AU - Imai, Hiroyuki
AU - Ono, Etsuro
N1 - Funding Information:
We appreciate the support from the Laboratory for technical support in the Medical Institute of Bioregulation, The Research Support Center, Research Center for Human Disease Modeling, Kyushu University Graduate School of Medical Sciences, and the Center for Advanced Instrumental and Educational Supports, Faculty of Agriculture (Kyushu University, Fukuoka, Japan). We thank E. Koba, M. Oda, K. Ichikawa, Y. Okugawa, and S. Takami for technical assistance; K. Tagawa and N. Takematsu as student laboratory assistants; and Prof. Douglas Drummond for critical review of the manuscript. We are grateful to Prof. Martin M. Matzuk, Baylor College of Medicine, US, for the TEX14 antibody, to Agilent Technologies for analyses of RNA, and to APRO Life Science Institute for analyses of proteomics data.
Funding Information:
∗Correspondence: Tokuko Iwamori, PhD, Department of Biomedicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan Center of Biomedical Research, Research Center for Human Disease Modeling, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi Higashi-ku, Fukuoka 812-8582, Japan. Tel: +81-92-642-6150; Fax: 81-92-642-6165, E-Mail: tiwamori@anim.med.kyushu-u.ac.jp †Grant Support: This work was supported in part by a Grant-in-Aid for Scientific Research (KAKENHI) on Innovative Areas “Mechanisms regulating gamete formation in animals” (26114506 to TI) and “Stem Cell Aging and Disease” (17H05648 to NI) from the Ministry of Education, Culture, Sports, Sciences, and Technology of Japan, Grant-in-Aid for Young Scientists (B) (15K21217, 17K15396 to TI), and (A) (26712026 to NI), Grant-in Aid for Scientific Research (B) (19H03139 to NI), and (C) (19K06440 to TI), Inamori Foundation (to TI), Takeda Science Foundation (to NI) and QR project of Kyushu University (2012KB6000 to TI). ‡Lead Author Conference Presentation: Presented in part at the 109th Annual Meeting of the Society of Reproduction and Development, 21–24 August 2017, Japan, and the 40th Annual Meeting of Biochemistry and Molecular Biology, 1–4 December 2017, Kobe Port Island, Hyogo, Japan.
Publisher Copyright:
© 2020 The Author(s) 2020. Published by Oxford University Press on behalf of Society for the Study of Reproduction. All rights reserved.
PY - 2020/4/24
Y1 - 2020/4/24
N2 - Intercellular bridges (ICBs) connecting germ cells are essential for spermatogenesis, and their deletion causes male infertility. However, the functions and component factors of ICBs are still unknown. We previously identified novel ICB-associated proteins by proteomics analysis using ICB enrichment. Here, we performed immunoprecipitation-proteomics analyses using antibodies specific to known ICB proteins MKLP1, RBM44, and ectoplasmic specialization-associated protein KIAA1210 and predicted protein complexes in the ICB cores. KIAA1210, its binding protein topoisomerase2B (TOP2B), and tight junction protein ZO1 were identified as novel ICB proteins. On the other hand, as well as KIAA1210 and TOP2B, MKLP1 and RBM44, but not TEX14, were localized at the XY body of spermatocytes, suggesting that there is a relationship between ICB proteins and meiotic chromosomes. Moreover, small RNAs interacted with an ICB protein complex that included KIAA1210, RBM44, and MKLP1. These results indicate dynamic movements of ICB proteins and suggest that ICB proteins could be involved not only in the communication between germ cells but also in their epigenetic regulation. Our results provide a novel perspective on the function of ICBs and could be helpful in revealing the biological function of the ICB.
AB - Intercellular bridges (ICBs) connecting germ cells are essential for spermatogenesis, and their deletion causes male infertility. However, the functions and component factors of ICBs are still unknown. We previously identified novel ICB-associated proteins by proteomics analysis using ICB enrichment. Here, we performed immunoprecipitation-proteomics analyses using antibodies specific to known ICB proteins MKLP1, RBM44, and ectoplasmic specialization-associated protein KIAA1210 and predicted protein complexes in the ICB cores. KIAA1210, its binding protein topoisomerase2B (TOP2B), and tight junction protein ZO1 were identified as novel ICB proteins. On the other hand, as well as KIAA1210 and TOP2B, MKLP1 and RBM44, but not TEX14, were localized at the XY body of spermatocytes, suggesting that there is a relationship between ICB proteins and meiotic chromosomes. Moreover, small RNAs interacted with an ICB protein complex that included KIAA1210, RBM44, and MKLP1. These results indicate dynamic movements of ICB proteins and suggest that ICB proteins could be involved not only in the communication between germ cells but also in their epigenetic regulation. Our results provide a novel perspective on the function of ICBs and could be helpful in revealing the biological function of the ICB.
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U2 - 10.1093/biolre/ioaa017
DO - 10.1093/biolre/ioaa017
M3 - Article
C2 - 31995159
AN - SCOPUS:85084167538
SN - 0006-3363
VL - 102
SP - 1134
EP - 1144
JO - Biology of reproduction
JF - Biology of reproduction
IS - 5
ER -