title: Mutagenesis of ARS2 Domains To Assess Possible Roles in Cell Cycle Progression and MicroRNA and Replication-Dependent Histone mRNA Biogenesis.
abstract: ARS2 is a regulator of RNA polymerase II transcript processing through its role in the maturation of distinct nuclear cap-binding complex (CBC)-controlled RNA families. In this study, we examined ARS2 domain function in transcript processing. Structural modeling based on the plant ARS2 orthologue, SERRATE, revealed 2 previously uncharacterized domains in mammalian ARS2: an N-terminal domain of unknown function (DUF3546), which is also present in SERRATE, and an RNA recognition motif (RRM) that is present in metazoan ARS2 but not in plants. Both the DUF3546 and zinc finger domain (ZnF) were required for association with microRNA and replication-dependent histone mRNA. Mutations in the ZnF disrupted interaction with FLASH, a key component in histone pre-mRNA processing. Mutations targeting the Mid domain implicated it in DROSHA interaction and microRNA biogenesis. The unstructured C terminus was required for interaction with the CBC protein CBP20, while the RRM was required for cell cycle progression and for binding to FLASH. Together, our results support a bridging model in which ARS2 plays a central role in RNA recognition and processing through multiple protein and RNA interactions.
authors: Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
authors: Connor
authors: C
authors: O'Sullivan
authors: Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
authors: Jennifer
authors: J
authors: Christie
authors: Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
authors: Marcus
authors: M
authors: Pienaar
authors: Department of Biology, University of Victoria, Victoria, BC, Canada.
authors: Jake
authors: J
authors: Gambling
authors: Department of Biology, University of Victoria, Victoria, BC, Canada.
authors: Philip E B
authors: PE
authors: Nickerson
authors: Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada.
authors: Spencer C
authors: SC
authors: Alford
authors: Department of Biology, University of Victoria, Victoria, BC, Canada.
authors: Robert L
authors: RL
authors: Chow
authors: Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada phoward@uvic.ca.
authors: Perry L
authors: PL
authors: Howard
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002453:Cell Cycle; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D006657:Histones / Q000235:genetics* / Q000378:metabolism; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D039502:Nuclear Cap-Binding Protein Complex; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D016196:S Phase; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D015854:Up-Regulation
chemical_list: D002135:Calcium-Binding Proteins; C119301:Casp8ap2 protein, mouse; D004268:DNA-Binding Proteins; D006657:Histones; D035683:MicroRNAs; D039502:Nuclear Cap-Binding Protein Complex; D009687:Nuclear Proteins; D012333:RNA, Messenger; C527300:Srrt protein, mouse; D014157:Transcription Factors