Effects of varying the spacing within the D,D-35-E motif in the catalytic region of retroviral integrase.
Virology
; 379(2): 223-33, 2008 Sep 30.
Article
em En
| MEDLINE
| ID: mdl-18687451
ABSTRACT
The catalytic domain of all retroviral integrases contains an Asp,Asp-35-Glu (D,D-35-E) motif with precisely 35 amino acids between the second aspartate and the glutamate. We have now made several mutations designed to alter the length or flexibility of a mobile loop within this 35-amino-acid spacer region in full-length Rous sarcoma virus integrase. Surprisingly, most of the mutants had enzymatic activity, including ones that shortened or lengthened the loop by up to 6 amino acids. Several size mutants exhibited the two biologically relevant activities of integrase in reactions with Mn(2+), although they were inactive with Mg(2+). No viruses containing integrase with an altered length, however, replicated in cell culture, and these viruses were blocked at the integration step. Thus, the conserved 35-amino-acid spacing is not absolutely required for enzymatic activity, but the correlation between infectivity and Mg(2+)-dependent activity supports magnesium as the metal cofactor used by integrase in vivo.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Integrases
/
Vírus do Sarcoma de Rous
Limite:
Animals
Idioma:
En
Revista:
Virology
Ano de publicação:
2008
Tipo de documento:
Article