TAILIEUCHUNG - Báo cáo khoa học: Regulation of ascidian Rel by its alternative splice variant

TheRel/NF-jBfamilyof transcription factorsplaykey roles in morphogenesis and immune responses. We reported previously that As-rel1 and As-rel2 of the ascidianHalo-cynthia roretziare involved in notochord formation. The As-rel1 protein is a typical Rel/NF-jB family member, whereas the As-rel2 protein is a novel truncated product of As-rel1 that lacks a nuclear localization signal and the unique C-terminal region. Here, we present conclusive evi-dence that As-rel1 and As-rel2 are generated from a single gene by alternative splicing | Eur. J. Biochem. 270 4459-4468 2003 FEBS 2003 doi Regulation of ascidian Rel by its alternative splice variant Narudo Kawai1 Masumi Shimada1 Hiroyuki Kawahara1 Noriyuki Satoh2 and Hideyoshi Yokosawa1 1 Department of Biochemistry Graduate School of Pharmaceutical Sciences Hokkaido University Sapporo 060-0812 Japan department of Zoology Graduate School of Science Kyoto University Kyoto 606-8502 Japan The Rel NF-KB family of transcription factors play key roles in morphogenesis and immune responses. We reported previously that As-rel1 and As-rel2 of the ascidian Halo-cynthia roretzi are involved in notochord formation. The As-rel1 protein is a typical Rel NF-KB family member whereas the As-rel2 protein is a novel truncated product of As-rel1 that lacks a nuclear localization signal and the unique C-terminal region. Here we present conclusive evidence that As-rel1 and As-rel2 are generated from a single gene by alternative splicing. We analyzed the roles of As-rel2 using cells transfected with As-rel1 or As-rel2 or both. As-rel1 was localized in the nucleus and As-rel2 in the cytoplasm when they were transfected individually. In contrast when they were transfected simultaneously both were localized in the nucleus because of the association of As-rel2 with As-rel1. In this case the transcriptional activity of As-rel1 was suppressed by As-rel2. Ascidian IkB was found to sequester As-rel1 in the cytoplasm and suppress its transcriptional activity when As-rel1 and IkB were transfected simultaneously. In contrast when As-rel1 and IkB were transfected together with As-rel2 As-rel1 was transported into the nucleus and its transcriptional activity was rescued from inhibition by IkB whereas As-rel2 remained localized in the cytoplasm suggesting IkB sequestration in the cytoplasm by As-rel2. From these findings we conclude that the alternative splice variant As-rel2 regulates the nuclear localization and transcriptional activity of As-rel1. .

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