Abstract

RNA editing alters pre-mRNA through site-selective adenosine dearnination, which results in codon changes that lead to the production of novel proteins. An enzyme that catalyzes this reaction, double-stranded RNA adenosine deaminase (ADARI), contains two N terminal Z-DNA-binding motifs, Z,,, and 4, the function of which is as yet unknown. In this study, multidimensional NMR spectroscopy was used to show that the topology of Z""""""""' is (X 10 1 (X2 (X3 P2 03. Long-range NOEs indicate that P1 and P3 interact with each other. Site-directed mutagenesis was used to identify residues in 0, 03 and the loop connecting 02 to 03 that affect Z-DNA binding. Also identified were I I hydrophobic residues that are essential for protein stability. Comparison with known structures reveals some similarity between Z,,, and (oc + 0) helix-tum-helix proteins, such as histone 5 and the family of hepatocyte nuclear factor-3 winged-helix-tum-helix transcription factors. Taken together, the structural and functional data suggest that recognition of Z-DNA by Z,,, involves residues in both the 0 helix and the C-terminal P-sheet.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR000995-27
Application #
6578162
Study Section
Project Start
2002-05-01
Project End
2003-04-30
Budget Start
Budget End
Support Year
27
Fiscal Year
2002
Total Cost
Indirect Cost

  Institution

Name
Massachusetts Institute of Technology
Department
Type
DUNS #
City
Cambridge
State
MA
Country
United States
Zip Code
02139

  Publications

Marintchev, Assen; Edmonds, Katherine A; Marintcheva, Boriana et al. (2009) Topology and regulation of the human eIF4A/4G/4H helicase complex in translation initiation. Cell 136:447-60
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Frueh, Dominique P; Leed, Alison; Arthanari, Haribabu et al. (2009) Time-shared HSQC-NOESY for accurate distance constraints measured at high-field in (15)N-(13)C-ILV methyl labeled proteins. J Biomol NMR 45:311-8
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Chen, Jingyang; Dupradeau, Francois-Yves; Case, David A et al. (2007) Nuclear magnetic resonance structural studies and molecular modeling of duplex DNA containing normal and 4'-oxidized abasic sites. Biochemistry 46:3096-107
Lentz, Margaret R; Kim, John P; Westmoreland, Susan V et al. (2005) Quantitative neuropathologic correlates of changes in ratio of N-acetylaspartate to creatine in macaque brain. Radiology 235:461-8
Kim, John P; Lentz, Margaret R; Westmoreland, Susan V et al. (2005) Relationships between astrogliosis and 1H MR spectroscopic measures of brain choline/creatine and myo-inositol/creatine in a primate model. AJNR Am J Neuroradiol 26:752-9
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