| dc.creator | VERA,JORGE | |
| dc.creator | VALENZUELA,BEATRIZ | |
| dc.creator | ROTH,MÓNICA J | |
| dc.creator | LEÓN,ÓSCAR | |
| dc.date | 2008-01-01 | |
| dc.date.accessioned | 2019-05-02T21:21:43Z | |
| dc.date.available | 2019-05-02T21:21:43Z | |
| dc.identifier | https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0716-97602008000100009 | |
| dc.identifier.uri | http://revistaschilenas.uchile.cl/handle/2250/81852 | |
| dc.description | Processing of viral DNA by retroviral integrase leaves a dinucleotide single-strand overhang in the unprocessed strand. Previous studies have stressed the importance of the 5' single-stranded (ss) tail in the integration process. To characterize the ss-tail binding site on M-MuLV integrase, we carried out crosslinking studies utilizing a disintegration substrate that mimics the covalent intermediate formed during integration. This substrate carried reactive groups at the 5' ss tail. A bromoacetyl derivative with a side chain of 6 A was crosslinked to the mutant IN 106-404, which lacks the N-terminal domain, yielding a crosslinked complex of 50 kDa. Treatment of IN 106-404 with N-ethylmaleimide (NEM) prevented crosslinking, suggesting that Cys209 was involved in the reaction. The reactivity of Cys209 was confirmed by crosslinking of a more specific derivative carrying maleimide groups that spans 8A approximately. In contrast, WT IN was not reactive, suggesting that the N-terminal domain modifies the reactivity of the Cys209 or the positioning of the crosslinker side chain. A similar oligonucleotide-carrying iodouridine at the 5'ss tail reacted with both IN 106-404 and WT IN upon UV irradiation. This reaction was also prevented by NEM, suggesting that the ss-tail positions near a peptide region that includes Cys209 | |
| dc.format | text/html | |
| dc.language | en | |
| dc.publisher | Sociedad de Biología de Chile | |
| dc.relation | 10.4067/S0716-97602008000100009 | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.source | Biological Research v.41 n.1 2008 | |
| dc.subject | integrase | |
| dc.subject | retrovirus | |
| dc.subject | crosslinking | |
| dc.title | Characterization of the long-terminal repeat single-strand tail-binding site of Moloney-MuLV integrase by crosslinking | |
