The circadian protein Nocturnin (NOCT) belongs to the exonuclease, endonuclease and phosphatase superfamily and is most similar to the CCR4-class of deadenylases that degrade the poly-adenosine tails of mRNAs. NOCT-deficient mice are resistant to high-fat diet induced weight gain, and exhibit dysregulation of bone formation. However, the mechanisms by which NOCT regulates these processes remain to be determined. Here, we describe a pair of high-resolution crystal structures of the human NOCT catalytic domain. The active site of NOCT is highly conserved with other exoribonucleases, and when directed to a transcript in cells, NOCT can reduce translation and abundance of that mRNA in a manner dependent on key active site residues. In contrast to the related deadenylase CNOT6L, purified recombinant NOCT lacks in vitro ribonuclease activity, suggesting that unidentified factors are necessary for enzymatic activity. We also find the ability of NOCT to repress reporter mRNAs in cells depends upon the 3′ end of the mRNA, as reporters terminating with a 3′ MALAT1 structure cannot be repressed by NOCT. Together, these data demonstrate that NOCT is an exoribonuclease that can degrade mRNAs to inhibit protein expression, suggesting a molecular mechanism for its regulatory role in lipid metabolism and bone development.
Bibliographical noteFunding Information:
Edward Mallinckrodt Jr. Foundation Grant (to A.C.G.); University of Michigan Nutrition and Obesity Research Center Pilot Grant (supported by the National Institutes of Health (NIH) grant [DK089503 to R.C.T.]; American Heart Association Predoctoral Fellowship [16PRE26700002 to E.T.A.]; NIH Chemistry-Biology Training Program Fellowship [5T32GM008597 to E.T.A.]; NIH Cellular Biotechnology Training Program Fellowship [5T32GM008353 to J.B.]; National Institute of General Medical Sciences, NIH grant [R01GM105707 to A.C.G.]; U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory [DE-AC02-06CH11357]; Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor [085P1000817]. Funding for open access charge: University of Minnesota, Institutional Funds. Conflict of interest statement. None declared.