title: N-TERMINALLY ELONGATED SpliInx2 AND SpliInx3 REDUCE BACULOVIRUS-TRIGGERED APOPTOSIS VIA HEMICHANNEL CLOSURE.
abstract: The hemichannel and gap junction channel are major portals for the release of factors responsible for the effects of apoptotic cells on the spread of apoptosis to neighboring cells and apoptotic corpse clearance, typically by phagocytes. The N-terminal cytoplasmic domain in the connexins, gap junction proteins in vertebrate, has been implicated in regulating channel closure. However, little is known about how the hemichannel close responds to apoptotic signaling transduction leading to the reduction of neighboring cellular apoptosis in an invertebrate. An insect Bac-to-Bac expression system, pFastBac(TM) HT A, allows us to construct an N-terminally elongated SpliInx2 (Nte-Inx2) and SpliInx3 (Nte-Inx3). Here, we demonstrated that recombinant baculovirus Bac-Nte-Inx2 (reBac-Net-Inx2) and Bac-Nte-Inx3 (reBac-Nte-Inx3) closed the endogenous hemichannel on the Sf9 cell surface. Importantly, primary baculovirus infections significantly caused early apoptosis, and this apoptosis was reduced by hemichannel-closed Sf9 cells at 24-h post-infection (PI). Although N-terminal-elongated residue led to the increase in the phosphorylated sites in both Nte-Inx2 and Nte-Inx3 and an additional transmembrane domain in Nte-Inx3, both the proteins localized on the cell surface, suggesting Nte-Inxs proteins could mediate hemichannel closure. Further supporting evidence showed that hemichannel closure was dependent on N-Inxs expressed by baculovirus polyhedrin promoter, which began to express at 18-24 h PI. These results identify an unconventional function of N-terminal-elongated innexins that could act as a plug to manipulate hemichannel closure and provide a mechanism connecting the effect of hemichannel closure directly to apoptotic signaling transduction from intracellular to extracellular compartment.
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Ya-Bin
authors.initials: YB
authors.lastname: Chen
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xiao
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Yang
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Jian-Sheng
authors.initials: JS
authors.lastname: Hu
authors.affiliation: School of Life Sciences, Yunnan University, Kunming, P. R. China.|Key Laboratory for Animal Genetic Diversity and Evolution of High Education in Yunnan Province, Yunnan University, Kunming, P. R. China.
authors.forename: Kai-Jun
authors.initials: KJ
authors.lastname: Luo
mesh_terms: D000818:Animals; D017209:Apoptosis; D016367:Baculoviridae / Q000502:physiology; D017630:Connexins / Q000235:genetics* / Q000378:metabolism; D017629:Gap Junctions / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000502:physiology / Q000821:virology; D015398:Signal Transduction; D018411:Spodoptera / Q000235:genetics / Q000254:growth & development / Q000502:physiology* / Q000821:virology
chemical_list: D017630:Connexins; D019476:Insect Proteins
keywords: N-terminus; apoptosis; baculovirus; hemichannel closure