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PMID: 14633971 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Structural and biochemical analyses of DNA and RNA binding by a bifunctional homing endonuclease and group I intron splicing factor.

Genes & development ·Vol. 17 ·No. 23 ·2003-12-01 ·Pages 2875-88

Bolduc JM, Spiegel PC, Chatterjee P, Brady KL, Downing ME, Caprara MG, Waring RB, Stoddard BL

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 A resolution. The structure demonstrates the remarkable structural conservation of the beta-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.

MeSH Terms
Amino Acid Sequence Base Sequence DNA/chemistry,metabolism Deoxyribonucleases, Type II Site-Specific/metabolism Introns Models, Molecular Molecular Sequence Data Nucleic Acid Conformation Protein Binding RNA/metabolism RNA Splicing Sequence Homology, Amino Acid
Chemicals
RNA DNA Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Bolduc Jill M
Fred Hutchinson Cancer Research Center, Division of Basic Sciences, Seattle, Washington 98109, USA.
Spiegel P Clint
Chatterjee Piyali
Brady Kristina L
Downing Maureen E
Caprara Mark G
Waring Richard B
Stoddard Barry L
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
0890-9369
DOI
10.1101/gad.1109003
Published
2003-12-01
Epub
2003-00-21
Pages
2875-88
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC289148
Subset
IM
Grants
NIGMS NIH HHS · R01 GM049857 · United States
NIGMS NIH HHS · R01 GM062853 · United States
NIGMS NIH HHS · GM49857 · United States
NIGMS NIH HHS · GM62853 · United States
Corrections
CommentIn
PMID: 14665667
External Links
PubMed source DOI: 10.1101/gad.1109003 PMC Full Text View journal details
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