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Abstract:
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X-ray diffraction data on a few retroviral integrases show a flexible loop near the active site. By sequence
alignment, the peptide region 207–218 of Mo-MLV IN appears to correspond to this flexible loop. In this
study, residues H208, Y211, R212, Q214, S215 and S216 of Mo-MLV IN were mutated to determine their
role on enzyme activity. We found that Y211A, R212A, R212K and Q214A decreased integration activity,
while disintegration and 30-processing were not significantly affected. By contrast H208A was completely
inactive in all the assays. The core domain of Mo-MLV integrase was modeled and the flexibility of the
region 207–216 was analyzed. Substitutions with low integration activity showed a lower flexibility than
wild type integrase. We propose that the peptide region 207–216 is a flexible loop and that H208, Y211,
R212 and Q214 of this loop are involved in the correct assembly of the DNA-integrase complex during
integration. |