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Titolo:
RETROVIRAL INTEGRATION - IN-VITRO HOST SITE SELECTION BY AVIAN INTEGRASE
Autore:
FITZGERALD ML; GRANDGENETT DP;
Indirizzi:
ST LOUIS UNIV,INST MOLEC VIROL,MED CTR,3681 PK AVE ST LOUIS MO 63110 ST LOUIS UNIV,INST MOLEC VIROL,MED CTR ST LOUIS MO 63110
Titolo Testata:
Journal of virology
fascicolo: 7, volume: 68, anno: 1994,
pagine: 4314 - 4321
SICI:
0022-538X(1994)68:7<4314:RI-IHS>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
MYELOBLASTOSIS VIRUS INTEGRASE; PROTEIN INVITRO; DNA INTEGRATION; RECOMBINATION; TERMINI;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
27
Recensione:
Indirizzi per estratti:
Citazione:
M.L. Fitzgerald e D.P. Grandgenett, "RETROVIRAL INTEGRATION - IN-VITRO HOST SITE SELECTION BY AVIAN INTEGRASE", Journal of virology, 68(7), 1994, pp. 4314-4321

Abstract

Viral integrase catalyzes the integration of the linear vir al DNA genome into the chromatin of the infected host cell, an essential step in the life cycle of retroviruses. The reaction produces a characteristic small duplication of host sequences at the site of integration, implying that there is a close juxtaposition of the viral DNA ends duringa concerted integration event. We have used an in vitro assay to measure the concerted integration of virus-like plasmid DNA into naked lambda DNA catalyzed by virion purified avian integrase. In contrast to in vivo avian integration, which has strong fidelity for a 6-bp duplication, purified avian integrase in the context of this assay produced adistribution of duplication sizes, with the 6-bp size dominating. Themetal cofactor Mg2+ induced increased fidelity for the 6-bp duplication relative to that with Mn2+. The immediate sequence of the host sitemay also influence duplication size in that we found sites that sustained multiple independent integration events producing the same duplication size. Additionally, for each set of cloned integration sites (5,6, and 7 bp), a unique but similar symmetrical pattern of GIC and AITsequence biases was found. Using duplex oligonucleotides as target substrates, we tested the significance of the 6-bp G/C and A/T pattern for site selection. In the context of this assay, which is likely dominated by the integration of only one viral end, the 6-bp pattern was not preferred. Instead, integration was predominately into the 3' ends of the oligonucleotides. The combined results of the lambda and oligonucleotide assays indicated that although host site selection has properties in common with recognition of the viral DNA termini, the nonrandom sequence preferences seen for host site selection were not identicalto the sequence requirements for long terminal repeat recognition.

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Documento generato il 28/05/20 alle ore 21:50:26