title: HMGA1a is involved in specific splice site regulation of human immunodeficiency virus type 1.
abstract: Human immunodeficiency virus type 1 (HIV-1) utilizes a highly complex splice site regulation system, taking advantage of host proteins, to express its own viral protein in an orderly way. We show here that one of the host proteins, high mobility group A protein 1a (HMGA1a), is involved in splice site regulation of 3' splice site 2 (A2) and 5'splice site 3 (D3) of HIV-1 genomic RNA. shRNA knockdown of HMGA1 in HeLa cells resulting in a decrease of HMGA1 showed a significant decrease of Vpr mRNA. RNA electrophoretic mobility shift assays showed HMGA1a specifically binds to a sequence adjacently upstream D3. In vitro splicing using heterologous pre-mRNA with A2 and D3, showed HMGA1a induced a splicing intermediate which decreased when an RNA decoy of the HMGA1a binding site was added. RT-PCR of in vitro splicing products revealed that HMGA1a induced an incomplete splicing product resulting from usage of A2 but inhibition of D3, which is reminiscent of the splicing pattern necessary for Vpr mRNA formation. HMGA1a interacted with hnRNPA1 shown by coimmunoprecipitation and supershifted U1 snRNP in an RNA electrophoretic mobility shift assay. We conclude that HMGA1a anchors U1 snRNP to inhibit D3 function, and that HMGA1a inhibits hnRNPA1 function on exon splicing silencer of Vpr (ESSV) to activate A2 function. We show here for the first time that HMGA1a is involved in specific splice site regulation of HIV-1.
authors.affiliation: National Institute of Infectious Diseases, Musashimurayama, Tokyo 208-0011, Japan.
authors.forename: Chikayuki
authors.initials: C
authors.lastname: Tsuruno
authors.forename: Kenji
authors.initials: K
authors.lastname: Ohe
authors.forename: Madoka
authors.initials: M
authors.lastname: Kuramitsu
authors.forename: Takuya
authors.initials: T
authors.lastname: Kohma
authors.forename: Youichi
authors.initials: Y
authors.lastname: Takahama
authors.forename: Yukio
authors.initials: Y
authors.lastname: Hamaguchi
authors.forename: Isao
authors.initials: I
authors.lastname: Hamaguchi
authors.forename: Kazu
authors.initials: K
authors.lastname: Okuma
mesh_terms: D017398:Alternative Splicing; D001483:Base Sequence; D055785:Gene Knockdown Techniques; D015497:HIV-1 / Q000235:genetics*; D025741:HMGA1a Protein / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006801:Humans; D022821:RNA Splice Sites; D012367:RNA, Viral / Q000235:genetics*
chemical_list: D022821:RNA Splice Sites; D012367:RNA, Viral; D025741:HMGA1a Protein