TY - JOUR
T1 - ESCRT Machinery Mediates Cytokinetic Abscission in the Unicellular Red Alga Cyanidioschyzon merolae
AU - Yagisawa, Fumi
AU - Fujiwara, Takayuki
AU - Takemura, Tokiaki
AU - Kobayashi, Yuki
AU - Sumiya, Nobuko
AU - Miyagishima, Shin Ya
AU - Nakamura, Soichi
AU - Imoto, Yuuta
AU - Misumi, Osami
AU - Tanaka, Kan
AU - Kuroiwa, Haruko
AU - Kuroiwa, Tsuneyoshi
N1 - Funding Information:
We thank Ms. K. Hashimoto and Y. Tanaka of the Miyagishima laboratory for technical support. We are grateful to Dr. Keiji Nishida (Kobe University), Dr. Mio Ohnuma (National Institute of technology, Hiroshima College), Dr. Takema Sasaki (National Institute of Genetics), and Dr. Yoshihisa Oda (National Institute of Genetics) for helpful advice during the study. We also thank the Biomaterials Analysis Division, Tokyo Institute of Technology, for DNA sequencing analysis. Fluorescence microscopy using the Keyence BZ-X700 was performed at the Center for Strategic Research Project of the University of the Ryukyus. We thank Mitchell Arico from Edanz Group for assistance in editing a draft of this manuscript. Funding. This study was supported by MEXT/JSPS KAKENHI 15H06520, 16K14770, and 19K06742 (to FY), 17K07439 (to YK), 18K06300 (to TF), and 19H03260 (to TK and FY), NIG-JOINT 2016-A1-4, 8A2017, 7A2018, and 31A2019 (to FY), and the University of the Ryukyus Research Project Promotion Grant for Women Researchers (to FY).
Publisher Copyright:
© Copyright © 2020 Yagisawa, Fujiwara, Takemura, Kobayashi, Sumiya, Miyagishima, Nakamura, Imoto, Misumi, Tanaka, Kuroiwa and Kuroiwa.
PY - 2020/4/3
Y1 - 2020/4/3
N2 - In many eukaryotes, cytokinesis proceeds in two successive steps: first, ingression of the cleavage furrow and second, abscission of the intercellular bridge. In animal cells, the actomyosin contractile ring is involved in the first step, while the endosomal sorting complex required for transport (ESCRT), which participates in various membrane fusion/fission events, mediates the second step. Intriguingly, in archaea, ESCRT is involved in cytokinesis, raising the hypothesis that the function of ESCRT in eukaryotic cytokinesis descended from the archaeal ancestor. In eukaryotes other than in animals, the roles of ESCRT in cytokinesis are poorly understood. To explore the primordial core mechanisms for eukaryotic cytokinesis, we investigated ESCRT functions in the unicellular red alga Cyanidioschyzon merolae that diverged early in eukaryotic evolution. C. merolae provides an excellent experimental system. The cell has a simple organelle composition. The genome (16.5 Mb, 5335 genes) has been completely sequenced, transformation methods are established, and the cell cycle is synchronized by a light and dark cycle. Similar to animal and fungal cells, C. merolae cells divide by furrowing at the division site followed by abscission of the intercellular bridge. However, they lack an actomyosin contractile ring. The proteins that comprise ESCRT-I–IV, the four subcomplexes of ESCRT, are partially conserved in C. merolae. Immunofluorescence of native or tagged proteins localized the homologs of the five ESCRT-III components [charged multivesicular body protein (CHMP) 1, 2, and 4–6], apoptosis-linked gene-2-interacting protein X (ALIX), the ESCRT-III adapter, and the main ESCRT-IV player vacuolar protein sorting (VPS) 4, to the intercellular bridge. In addition, ALIX was enriched around the cleavage furrow early in cytokinesis. When the ESCRT function was perturbed by expressing dominant-negative VPS4, cells with an elongated intercellular bridge accumulated—a phenotype resulting from abscission failure. Our results show that ESCRT mediates cytokinetic abscission in C. merolae. The fact that ESCRT plays a role in cytokinesis in archaea, animals, and early diverged alga C. merolae supports the hypothesis that the function of ESCRT in cytokinesis descended from archaea to a common ancestor of eukaryotes.
AB - In many eukaryotes, cytokinesis proceeds in two successive steps: first, ingression of the cleavage furrow and second, abscission of the intercellular bridge. In animal cells, the actomyosin contractile ring is involved in the first step, while the endosomal sorting complex required for transport (ESCRT), which participates in various membrane fusion/fission events, mediates the second step. Intriguingly, in archaea, ESCRT is involved in cytokinesis, raising the hypothesis that the function of ESCRT in eukaryotic cytokinesis descended from the archaeal ancestor. In eukaryotes other than in animals, the roles of ESCRT in cytokinesis are poorly understood. To explore the primordial core mechanisms for eukaryotic cytokinesis, we investigated ESCRT functions in the unicellular red alga Cyanidioschyzon merolae that diverged early in eukaryotic evolution. C. merolae provides an excellent experimental system. The cell has a simple organelle composition. The genome (16.5 Mb, 5335 genes) has been completely sequenced, transformation methods are established, and the cell cycle is synchronized by a light and dark cycle. Similar to animal and fungal cells, C. merolae cells divide by furrowing at the division site followed by abscission of the intercellular bridge. However, they lack an actomyosin contractile ring. The proteins that comprise ESCRT-I–IV, the four subcomplexes of ESCRT, are partially conserved in C. merolae. Immunofluorescence of native or tagged proteins localized the homologs of the five ESCRT-III components [charged multivesicular body protein (CHMP) 1, 2, and 4–6], apoptosis-linked gene-2-interacting protein X (ALIX), the ESCRT-III adapter, and the main ESCRT-IV player vacuolar protein sorting (VPS) 4, to the intercellular bridge. In addition, ALIX was enriched around the cleavage furrow early in cytokinesis. When the ESCRT function was perturbed by expressing dominant-negative VPS4, cells with an elongated intercellular bridge accumulated—a phenotype resulting from abscission failure. Our results show that ESCRT mediates cytokinetic abscission in C. merolae. The fact that ESCRT plays a role in cytokinesis in archaea, animals, and early diverged alga C. merolae supports the hypothesis that the function of ESCRT in cytokinesis descended from archaea to a common ancestor of eukaryotes.
KW - Cyanidioschyzon merolae
KW - ESCRT
KW - cytokinesis
KW - cytokinetic abscission
KW - red alga
UR - http://www.scopus.com/inward/record.url?scp=85083509516&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85083509516&partnerID=8YFLogxK
U2 - 10.3389/fcell.2020.00169
DO - 10.3389/fcell.2020.00169
M3 - Article
AN - SCOPUS:85083509516
SN - 2296-634X
VL - 8
JO - Frontiers in Cell and Developmental Biology
JF - Frontiers in Cell and Developmental Biology
M1 - 169
ER -