The multimerization state of retroviral RNA is modulated by ammonium ions and affects HIV-1 full-length cDNA synthesis in vitro.
Genomic human immunodeficiency virus type 1 (HIV-1) RNA fragments containing the dimer linkage structure (DLS) can be dimerized and multimerized in the presence of NH4+ and in the absence of any other cation and any viral or cellular protein. This effect strongly supports the notion that dimerization and multimerization of genomic RNA occurs via purine-quartet formation in quadruple helical RNA structures. The efficiency of RNA dimerization and multimerization in the presence of ammonium ions is about 400 fold increased as compared to alkali metal ions such as potassium. Dimerized retroviral RNA representing a pseudodiploid genome could account for genetic recombination within the virion and during reverse transcription. Application of a novel South-Northern-Blotting procedure with biotinylated RNA and digoxigenin-labelled cDNA in vitro reveals that efficient human- and bovine tRNA(Lys3) primed full-length cDNA-synthesis only takes place with a predominantly monomerized RNA template. Dimerization and multimerization of the RNA significantly reduces full-length cDNA-synthesis. This suggests that monomerization of the dimerized RNA, effected by deionization in vitro, is essential for efficient retroviral reverse transcription in vivo.
Weiss
S
S
Institut für Biochemie, Universität München, Germany.
Häusl
G
G
Famulok
M
M
König
B
B
D000085:Acetates / Q000737:chemistry; D001704:Biopolymers / Q000737:chemistry; D001710:Biotin; D015152:Blotting, Northern / Q000379:methods; D015139:Blotting, Southern / Q000379:methods; D004279:DNA, Viral / Q000096:biosynthesis* / Q000737:chemistry; D004076:Digoxigenin; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000235:genetics*; D007477:Ions; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry*; D012190:Retroviridae / Q000235:genetics*
D000085:Acetates; D001704:Biopolymers; D004279:DNA, Viral; D007477:Ions; D000644:Quaternary Ammonium Compounds; D012367:RNA, Viral; D001710:Biotin; D004076:Digoxigenin; C018824:ammonium acetate