The disordered C-terminal domain of human DNA glycosylase NEIL1 contributes to its stability via intramolecular interactions

Muralidhar L. Hegde, Susan E. Tsutakawa, Pavana M. Hegde, Luis Marcelo F. Holthauzen, Jing Li, Numan Oezguen, Vincent J. Hilser, John A. Tainer, Sankar Mitra

Research output: Contribution to journalArticlepeer-review

38 Scopus citations


NEIL1 [Nei (endonuclease VIII)-like protein 1], one of the five mammalian DNA glycosylases that excise oxidized DNA base lesions in the human genome to initiate base excision repair, contains an intrinsically disordered C-terminal domain (CTD; ∼ 100 residues), not conserved in its Escherichia coli prototype Nei. Although dispensable for NEIL1's lesion excision and AP lyase activities, this segment is required for efficient in vivo enzymatic activity and may provide an interaction interface for many of NEIL1's interactions with other base excision repair proteins. Here, we show that the CTD interacts with the folded domain in native NEIL1 containing 389 residues. The CTD is poised for local folding in an ordered structure that is induced in the purified fragment by osmolytes. Furthermore, deletion of the disordered tail lacking both Tyr and Trp residues causes a red shift in NEIL1's intrinsic Trp-specific fluorescence, indicating a more solvent-exposed environment for the Trp residues in the truncated protein, which also exhibits reduced stability compared to the native enzyme. These observations are consistent with stabilization of the native NEIL1 structure via intramolecular, mostly electrostatic, interactions that were disrupted by mutating a positively charged (Lys-rich) cluster of residues (amino acids 355-360) near the C-terminus. Small-angle X-ray scattering (SAXS) analysis confirms the flexibility and dynamic nature of NEIL1's CTD, a feature that may be critical to providing specificity for NEIL1's multiple, functional interactions.

Original languageEnglish (US)
Pages (from-to)2359-2371
Number of pages13
JournalJournal of Molecular Biology
Issue number13
StatePublished - Jul 10 2013


  • DNA repair
  • electrostatic interactions
  • intrinsically unstructured region
  • oxidative DNA damage
  • protein stability

ASJC Scopus subject areas

  • Molecular Biology


Dive into the research topics of 'The disordered C-terminal domain of human DNA glycosylase NEIL1 contributes to its stability via intramolecular interactions'. Together they form a unique fingerprint.

Cite this