Document Type
Article
Publication Date
2003
Keywords
Homing endonuclease, Maturase, Group I intron, Crystal structure, DNA binding
Abstract
We determined the crystal structure of a bifunctional group I intron splicing factor and homing endonuclease, termed the I-AniI maturase, in complex with its DNA target at 2.6 Å resolution. The structure demonstrates the remarkable structural conservation of the (3-sheet DNA-binding motif between highly divergent enzyme subfamilies. DNA recognition by I-AniI was further studied using nucleoside deletion and DMS modification interference analyses. Correlation of these results with the crystal structure provides information on the relative importance of individual nucleotide contacts for DNA recognition. Alignment and modeling of two homologous maturases reveals conserved basic surface residues, distant from the DNA-binding surface, that might be involved in RNA binding. A point mutation that introduces a single negative charge in this region uncouples the maturase and endonuclease functions of the protein, inhibiting RNA binding and splicing while maintaining DNA binding and cleavage.
Publication Title
Genes & Development
Volume
17
Issue
23
First Page
2875
Last Page
2888
Required Publisher's Statement
Published by Cold Spring Harbor Laboratory Press
DOI: 10.1101/gad.1109003
Recommended Citation
Bolduc, Jill M.; Spiegel, P. Clint; Chatterjee, Pivali; Brady, Kristina L.; Downing, Maureen E.; Caprara, Mark G.; Waring, Richard B.; and Stoddard, Barry L., "Structural and Biochemical Analyses of DNA and RNA Binding by a Bifunctional Homing Endonuclease and Group I Intron Splicing Factor" (2003). Chemistry Faculty and Staff Publications. 6.
https://cedar.wwu.edu/chemistry_facpubs/6
Subjects - Topical (LCSH)
Endonucleases; DNA-binding proteins--Analysis; RNA splicing--Analysis
Genre/Form
articles
Type
Text
Language
English
Format
application/pdf
