Abstract
A Nudix enzyme from Bacillus cereus (NCBI RefSeq accession no. NP_831800) catalyzes the hydrolysis of CDP-choline to produce CMP and phosphocholine. Here, we show that in addition, the enzyme has a 3'→5' RNA exonuclease activity. The structure of the free enzyme, determined to a 1.8-Å resolution, shows that the enzyme is an asymmetric dimer. Each monomer consists of two domains, an N-terminal helical domain and a C-terminal Nudix domain. The N-terminal domain is placed relative to the C-terminal domain such as to result in an overall asymmetric arrangement with two distinct catalytic sites: one with an "enclosed" Nudix pyrophosphatase site and the other with a more open, less-defined cavity. Residues that may be important for determining the asymmetry are conserved among a group of uncharacterized Nudix enzymes from Gram-positive bacteria. Our data support a model where CDP-choline hydrolysis is catalyzed by the enclosed Nudix site and RNA exonuclease activity is catalyzed by the open site. CDPChase is the first identified member of a novel Nudix family in which structural asymmetry has a profound effect on the recognition of substrates.
Original language | English (US) |
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Pages (from-to) | 3175-3185 |
Number of pages | 11 |
Journal | Journal of bacteriology |
Volume | 193 |
Issue number | 13 |
DOIs | |
State | Published - Jul 2011 |
Externally published | Yes |
ASJC Scopus subject areas
- Microbiology
- Molecular Biology