Other search tools

About this data

The publication data currently available has been vetted by Vanderbilt faculty, staff, administrators and trainees. The data itself is retrieved directly from NCBI's PubMed and is automatically updated on a weekly basis to ensure accuracy and completeness.

If you have any questions or comments, please contact us.

Results: 1 to 3 of 3

Publication Record

Connections

The bacterial ribonuclease P holoenzyme requires specific, conserved residues for efficient catalysis and substrate positioning.
Reiter NJ, Osterman AK, Mondragón A
(2012) Nucleic Acids Res 40: 10384-93
MeSH Terms: Bacterial Proteins, Biocatalysis, Catalytic Domain, Models, Molecular, Mutagenesis, Site-Directed, RNA Precursors, RNA, Transfer, Ribonuclease P, Thermotoga maritima
Show Abstract · Added January 28, 2014
RNase P is an RNA-based enzyme primarily responsible for 5'-end pre-tRNA processing. A structure of the bacterial RNase P holoenzyme in complex with tRNAPhe revealed the structural basis for substrate recognition, identified the active site location, and showed how the protein component increases functionality. The active site includes at least two metal ions, a universal uridine (U52), and P RNA backbone moieties, but it is unclear whether an adjacent, bacterially conserved protein loop (residues 52-57) participates in catalysis. Here, mutagenesis combined with single-turnover reaction kinetics demonstrate that point mutations in this loop have either no or modest effects on catalytic efficiency. Similarly, amino acid changes in the 'RNR' region, which represent the most conserved region of bacterial RNase P proteins, exhibit negligible changes in catalytic efficiency. However, U52 and two bacterially conserved protein residues (F17 and R89) are essential for efficient Thermotoga maritima RNase P activity. The U52 nucleotide binds a metal ion at the active site, whereas F17 and R89 are positioned >20 Å from the cleavage site, probably making contacts with N(-4) and N(-5) nucleotides of the pre-tRNA 5'-leader. This suggests a synergistic coupling between transition state formation and substrate positioning via interactions with the leader.
0 Communities
1 Members
0 Resources
9 MeSH Terms
Structure of a bacterial ribonuclease P holoenzyme in complex with tRNA.
Reiter NJ, Osterman A, Torres-Larios A, Swinger KK, Pan T, Mondragón A
(2010) Nature 468: 784-9
MeSH Terms: Biocatalysis, Catalytic Domain, Crystallography, X-Ray, Genes, Bacterial, Holoenzymes, Metals, Models, Molecular, Molecular Conformation, RNA, Transfer, Phe, Ribonuclease P, Structure-Activity Relationship, Substrate Specificity, Thermotoga maritima
Show Abstract · Added January 28, 2014
Ribonuclease (RNase) P is the universal ribozyme responsible for 5'-end tRNA processing. We report the crystal structure of the Thermotoga maritima RNase P holoenzyme in complex with tRNA(Phe). The 154 kDa complex consists of a large catalytic RNA (P RNA), a small protein cofactor and a mature tRNA. The structure shows that RNA-RNA recognition occurs through shape complementarity, specific intermolecular contacts and base-pairing interactions. Soaks with a pre-tRNA 5' leader sequence with and without metal help to identify the 5' substrate path and potential catalytic metal ions. The protein binds on top of a universally conserved structural module in P RNA and interacts with the leader, but not with the mature tRNA. The active site is composed of phosphate backbone moieties, a universally conserved uridine nucleobase, and at least two catalytically important metal ions. The active site structure and conserved RNase P-tRNA contacts suggest a universal mechanism of catalysis by RNase P.
0 Communities
1 Members
0 Resources
13 MeSH Terms
Structure of the C-terminal half of UvrC reveals an RNase H endonuclease domain with an Argonaute-like catalytic triad.
Karakas E, Truglio JJ, Croteau D, Rhau B, Wang L, Van Houten B, Kisker C
(2007) EMBO J 26: 613-22
MeSH Terms: Amino Acid Motifs, Bacterial Proteins, Catalytic Domain, DNA-Binding Proteins, Endodeoxyribonucleases, Magnesium, Manganese, Models, Molecular, Protein Structure, Tertiary, Thermotoga maritima
Show Abstract · Added April 3, 2018
Removal and repair of DNA damage by the nucleotide excision repair pathway requires two sequential incision reactions, which are achieved by the endonuclease UvrC in eubacteria. Here, we describe the crystal structure of the C-terminal half of UvrC, which contains the catalytic domain responsible for 5' incision and a helix-hairpin-helix-domain that is implicated in DNA binding. Surprisingly, the 5' catalytic domain shares structural homology with RNase H despite the lack of sequence homology and contains an uncommon DDH triad. The structure also reveals two highly conserved patches on the surface of the protein, which are not related to the active site. Mutations of residues in one of these patches led to the inability of the enzyme to bind DNA and severely compromised both incision reactions. Based on our results, we suggest a model of how UvrC forms a productive protein-DNA complex to excise the damage from DNA.
0 Communities
1 Members
0 Resources
MeSH Terms