Effect of dimerization on the conformation of the encapsidation Psi domain of Moloney murine leukemia virus RNA.
In Moloney murine leukemia virus, the encapsidation Psi element was shown to be necessary and sufficient to promote packaging of viral RNA, and to be required for dimerization. The conformation of the Psi domain (nucleotides 215 to 565) was investigated in solution by chemical probing. The four bases were monitored at one of their Watson-Crick positions with dimethylsulfate at cytosine N3 and adenosine N1, and with a carbodiimide derivative at guanosine N1 and uridine N3. Position N7 of adenine residues was probed with diethylpyrocarbonate. The analyses were conducted on in vitro transcribed fragments corresponding either to the isolated Psi domain or to the 5'-terminal 725 nucleotides. The RNA fragments were analyzed in their monomeric and dimeric forms. A secondary structure model was derived from probing data, computer prediction and sequence analysis of related murine retroviruses. One major result is that Psi forms an independent and highly structured domain. Dimerization induces an extensive reduction of reactivity in region 278 to 309 that can be interpreted as the result of intermolecular interactions and/or intramolecular conformational rearrangements. A second region (around position 215) was shown to display discrete reactivity changes upon dimerization. These two regions represent likely elements of dimerization. More unexpectedly, reactivity changes (essentially enhancement of reactivity) were also detected in another part of Psi (around position 480) not believed to contain elements of dimerization. These reactivity changes could be interpreted as dimerization-induced allosteric transitions.
Tounekti
N
N
Unité de Biochimie, URA 158 CNRS, Institut Gustave Roussy, Villejuif, France.
Mougel
M
M
Roy
C
C
Marquet
R
R
Darlix
J L
JL
Paoletti
J
J
Ehresmann
B
B
Ehresmann
C
C
D001483:Base Sequence; D006860:Hydrogen Bonding; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008979:Moloney murine leukemia virus / Q000648:ultrastructure*; D009690:Nucleic Acid Conformation; D010802:Phylogeny; D012367:RNA, Viral / Q000737:chemistry / Q000648:ultrastructure*; D016415:Sequence Alignment
D046911:Macromolecular Substances; D012367:RNA, Viral