Two short basic sequences surrounding the zinc finger of nucleocapsid protein NCp10 of Moloney murine leukemia virus are critical for RNA annealing activity.
The 56 amino acid nucleocapsid protein (NCp10) of Moloney Murine Leukemia Virus, contains a CysX2CysX4HisX4Cys zinc finger flanked by basic residues. In vitro NCp10 promotes genomic RNA dimerization, a process most probably linked to genomic RNA packaging, and replication primer tRNA(Pro) annealing to the initiation site of reverse transcription. To characterize the amino-acid sequences involved in the various functions of NCp10, we have synthesized by solid phase method the native protein and a series of derived peptides shortened at the N- or C-terminus with or without the zinc finger domain. In the latter case, the two parts of the protein were linked by a Glycine - Glycine spacer. The in vitro studies of these peptides show that nucleic acid annealing activities of NCp10 do not require a zinc finger but are critically dependent on the presence of specific sequences located on each side of the CCHC domain and containing proline and basic residues. Thus, deletion of 11R or 49PRPQT, of the fully active 29 residue peptide 11RQGGERRRSQLDRDGGKKPRGPRGPRPQT53 leads to a complete loss of NCp10 activity. Therefore it is proposed that in NCp10, the zinc finger directs the spatial recognition of the target RNAs by the basic domains surrounding the zinc finger.
De Rocquigny
H
H
Unite de Pharmacochimie moléculaire et structurale, U266 INSERM, URA D1500 CNRS, Université René Descartes, Paris, France.
Ficheux
D
D
Gabus
C
C
Allain
B
B
Fournie-Zaluski
M C
MC
Darlix
J L
JL
Roques
B P
BP
D000595:Amino Acid Sequence; D015683:Gene Products, gag / Q000737:chemistry / Q000502:physiology*; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000235:genetics*; D010446:Peptide Fragments; D011392:Proline; D012361:RNA, Transfer, Pro / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D013329:Structure-Activity Relationship; D014158:Transcription, Genetic / Q000502:physiology*; D014758:Viral Core Proteins / Q000737:chemistry / Q000502:physiology*; D016335:Zinc Fingers / Q000502:physiology
D015683:Gene Products, gag; D010446:Peptide Fragments; D012361:RNA, Transfer, Pro; D012367:RNA, Viral; D014758:Viral Core Proteins; C068662:retroviral nucleocapsid protein NCp10, Moloney murine leukemia virus; D011392:Proline