Unknown

Dataset Information

0

Transition-state stabilization in Escherichia coli ribonuclease P RNA-mediated cleavage of model substrates.


ABSTRACT: We have used model substrates carrying modified nucleotides at the site immediately 5' of the canonical RNase P cleavage site, the -1 position, to study Escherichia coli RNase P RNA-mediated cleavage. We show that the nucleobase at -1 is not essential but its presence and identity contribute to efficiency, fidelity of cleavage and stabilization of the transition state. When U or C is present at -1, the carbonyl oxygen at C2 on the nucleobase contributes to transition-state stabilization, and thus acts as a positive determinant. For substrates with purines at -1, an exocyclic amine at C2 on the nucleobase promotes cleavage at an alternative site and it has a negative impact on cleavage at the canonical site. We also provide new insights into the interaction between E. coli RNase P RNA and the -1 residue in the substrate. Our findings will be discussed using a model where bacterial RNase P cleavage proceeds through a conformational-assisted mechanism that positions the metal(II)-activated H2O for an in-line attack on the phosphorous atom that leads to breakage of the phosphodiester bond.

SUBMITTER: Wu S 

PROVIDER: S-EPMC3874170 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

altmetric image

Publications

Transition-state stabilization in Escherichia coli ribonuclease P RNA-mediated cleavage of model substrates.

Wu Shiying S   Chen Yu Y   Mao Guanzhong G   Trobro Stefan S   Kwiatkowski Marek M   Kirsebom Leif A LA  

Nucleic acids research 20131003 1


We have used model substrates carrying modified nucleotides at the site immediately 5' of the canonical RNase P cleavage site, the -1 position, to study Escherichia coli RNase P RNA-mediated cleavage. We show that the nucleobase at -1 is not essential but its presence and identity contribute to efficiency, fidelity of cleavage and stabilization of the transition state. When U or C is present at -1, the carbonyl oxygen at C2 on the nucleobase contributes to transition-state stabilization, and thu  ...[more]

Similar Datasets

| S-EPMC10465520 | biostudies-literature
| S-EPMC2373462 | biostudies-literature
| S-EPMC4666806 | biostudies-literature
| S-EPMC3867540 | biostudies-literature
| S-EPMC7544649 | biostudies-literature
| S-EPMC7380570 | biostudies-literature
| S-EPMC2720131 | biostudies-literature
| S-EPMC3740856 | biostudies-literature
| S-EPMC2935283 | biostudies-literature
| S-EPMC2271172 | biostudies-literature