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J Mol Biol. 2000 Jul 21;300(4):867-75.

A model for the PI-SceIxDNA complex based on multiple base and phosphate backbone-specific photocross-links.

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1
Institut für Biochemie Fachbereich 08, Justus-Liebig-Universität, Heinrich-Buff-Ring 58, Giessen, D-35392, Germany.

Abstract

We have synthesized different oligodeoxynucleotides carrying, in single positions of the >36 bp recognition site of PI-SceI, photoreactive base analogues (5-iododeoxypyrimidines) or phosphate modifications (p-azidophenacylphosphorothioates) and used them in photocross-linking experiments with PI-SceI to probe the protein-DNA interface of the specific complex between the homing endonuclease PI-SceI and its DNA substrate. One base-specific and several backbone-specific cross-links were analyzed in detail: the cross-linking positions were identified by Edman degradation of isolated cross-linked peptidexoligodeoxynucleotide adducts and confirmed by site-directed mutagenesis. Based on these results and the crystal structure of PI-SceI, a model for the structure of the PI-SceIxDNA complex is proposed.

PMID:
10891273
DOI:
10.1006/jmbi.2000.3872
[Indexed for MEDLINE]
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