Medizinische Universität Graz - Research portal
Selected Publication:
SHR Neuro Cancer Cardio Lipid Metab Microb
Oliva, C; Sánchez-Murcia, PA; Rico, E; Bravo, A; Menéndez, M; Gago, F; Jiménez-Ruiz, A.
Structure-based domain assignment in Leishmania infantum EndoG: characterization of a pH-dependent regulatory switch and a C-terminal extension that largely dictates DNA substrate preferences.
Nucleic Acids Res. 2017; 45(15):9030-9045
Doi: 10.1093/nar/gkx629
[OPEN ACCESS]
Web of Science
PubMed
FullText
FullText_MUG
- Co-authors Med Uni Graz
- Sánchez Murcia Pedro Alejandro
- Altmetrics:
- Dimensions Citations:
- Plum Analytics:
- Scite (citation analytics):
- Abstract:
- Mitochondrial endonuclease G from Leishmania infantum (LiEndoG) participates in the degradation of double-stranded DNA (dsDNA) during parasite cell death and is catalytically inactive at a pH of 8.0 or above. The presence, in the primary sequence, of an acidic amino acid-rich insertion exclusive to trypanosomatids and its spatial position in a homology-built model of LiEndoG led us to postulate that this peptide stretch might act as a pH sensor for self-inhibition. We found that a LiEndoG variant lacking residues 145-180 is indeed far more active than its wild-type counterpart at pH values >7.0. In addition, we discovered that (i) LiEndoG exists as a homodimer, (ii) replacement of Ser211 in the active-site SRGH motif with the canonical aspartate from the DRGH motif of other nucleases leads to a catalytically deficient enzyme, (iii) the activity of the S211D variant can be restored upon the concomitant replacement of Ala247 with Arg and (iv) a C-terminal extension is responsible for the observed preferential cleavage of single-stranded DNA (ssDNA) and ssDNA-dsDNA junctions. Taken together, our results support the view that LiEndoG is a multidomain molecular machine whose nuclease activity can be subtly modulated or even abrogated through architectural changes brought about by environmental conditions and interaction with other binding partners. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- Amino Acid Substitution -
- Catalytic Domain -
- Cloning, Molecular -
- DNA Cleavage -
- DNA, Protozoan - chemistry
- DNA, Protozoan - genetics
- DNA, Protozoan - metabolism
- DNA, Single-Stranded - chemistry
- DNA, Single-Stranded - genetics
- DNA, Single-Stranded - metabolism
- Endodeoxyribonucleases - chemistry
- Endodeoxyribonucleases - genetics
- Endodeoxyribonucleases - metabolism
- Escherichia coli - genetics
- Escherichia coli - metabolism
- Gene Expression -
- Hydrogen-Ion Concentration -
- Kinetics -
- Leishmania infantum - chemistry
- Leishmania infantum - enzymology
- Models, Molecular -
- Nucleic Acid Conformation -
- Protein Binding -
- Protein Conformation, alpha-Helical -
- Protein Conformation, beta-Strand -
- Protein Interaction Domains and Motifs -
- Protein Multimerization -
- Protozoan Proteins - chemistry
- Protozoan Proteins - genetics
- Protozoan Proteins - metabolism
- Recombinant Proteins - chemistry
- Recombinant Proteins - genetics
- Recombinant Proteins - metabolism
- Sequence Alignment -
- Sequence Deletion -
- Sequence Homology, Amino Acid -
- Structure-Activity Relationship -
- Substrate Specificity -