HIV-1 Gag binds specifically to RNA stem-loops in the 5' leader sequence.
Biochim Biophys Acta
; 1398(3): 305-14, 1998 Jul 09.
Article
in En
| MEDLINE
| ID: mdl-9655923
ABSTRACT
GST-Gag(p55) binds specifically to HIV-1 RNA sequences 1-406, in vitro, with a Kd of about 50 nM. This RNA transcript contains a number of stem loop (SL) structures. The binding is due to the Gag moiety of the fusion protein, not GST. There is a high affinity binding site for Gag in an RNA containing nucleotides 325-362. SL4 is predicted by both biochemical studies and computer folding to be located between nucleotides 335 and 358. An RNA transcript ending at nucleotide 335 does not bind Gag. The deletion of nucleotides 334-358 from HIV-1 RNAs does not affect Gag binding. Digestions with RNase V1 and T1 show that nucleotides 297-300 in SL2, 310, 312, 313, 315, 317, 318, 325 in SL3, and 342 and 343 in SL4 are protected in the presence of Gag. The cleavage of nucleotides 348-351 in SL4 by RNAse V1 is enhanced by Gag binding. At least two Gag binding sites are therefore located in the leader RNA. Those located 5' of nucleotide 335 require the presence of additional 3' sequences.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Precursors
/
RNA, Messenger
/
RNA, Viral
/
Gene Products, gag
/
HIV-1
Limits:
Humans
Language:
En
Journal:
Biochim Biophys Acta
Year:
1998
Type:
Article
Affiliation country:
United kingdom