title: The length of an internal poly(A) tract of hibiscus latent Singapore virus is crucial for its replication.
abstract: Hibiscus latent Singapore virus (HLSV) mutants were constructed to study roles of its internal poly(A) tract (IPAT) in viral replication and coat protein (CP) expression. Shortening of the IPAT resulted in reduced HLSV RNA accumulation and its minimal length required for HLSV CP expression in plants was 24 nt. Disruption of a putative long range RNA-RNA interacting structure between 5' and 3' untranslated regions of HLSV-22A and -24A resulted in reduced viral RNA and undetectable CP accumulation in inoculated leaves. Replacement of the IPAT in HLSV with an upstream pseudoknot domain (UPD) of Tobacco mosaic virus (TMV) or insertion of the UPD to the immediate downstream of a 24 nt IPAT in HLSV resulted in drastically reduced viral RNA replication. Plants infected with a TMV mutant by replacement of the UPD with 43 nt IPAT exhibited milder mosaic symptoms without necrosis. We have proposed a model for HLSV replication.
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543; Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Science, Haikou, Hainan, China 571101.
authors.forename: Shengniao
authors.initials: S
authors.lastname: Niu
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Shishu
authors.initials: S
authors.lastname: Cao
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Li-Jing
authors.initials: LJ
authors.lastname: Huang
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Kelvin Chee-Leong
authors.initials: KC
authors.lastname: Tan
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543; Temasek Life Sciences Laboratory, 1 Research Link, Singapore 117604; National University of Singapore Suzhou Research Institute, Suzhou, Jiangsu, China 215123. Electronic address: dbswsm@nus.edu.sg.
authors.forename: Sek-Man
authors.initials: SM
authors.lastname: Wong
mesh_terms: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D001483:Base Sequence; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D016679:Genome, Viral; D031584:Hibiscus / Q000821:virology*; D008969:Molecular Sequence Data; D009154:Mutation; D009690:Nucleic Acid Conformation; D010935:Plant Diseases / Q000821:virology; D039102:Poly(A)-Binding Proteins / Q000235:genetics / Q000378:metabolism; D020871:RNA Stability; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014026:Nicotiana / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000235:genetics; D017898:Tobamovirus / Q000235:genetics* / Q000502:physiology* / Q000648:ultrastructure; D014771:Virion / Q000648:ultrastructure; D014779:Virus Replication / Q000235:genetics / Q000502:physiology
chemical_list: D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D036022:Capsid Proteins; D039102:Poly(A)-Binding Proteins; D012333:RNA, Messenger; D012367:RNA, Viral
keywords: CP expression; Internal poly(A) tract; Poly(A) binding protein; RNA kissing structure; RNA stability; Replication; Tobamovirus