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Certain adenosine residues within mammalian RNAs undergo reversible N6 methylation. Two methyltransferase enzymes, METTL3 and METTL14, as well as the splicing factor WTAP are identified as core components of the multiprotein complex that deposits RNA N6-methyladenosine (m6A) in nuclear RNAs.
Crystal structures reveal that the antitubercular compound pyridomycin blocks binding of the NADH cofactor to the fatty acid synthesis enzyme InhA while also blocking the lipid substrate–binding pocket.
Partial agonists are generally thought to promote GPCR conformations that signal suboptimally. Analysis of bifunctional muscarinic M2 receptor ligands now shows that partial agonism can also be predictably defined by a single ligand binding two receptor populations in different orientations.
Aspartate is a primary nitrogen source for Mycobacterium tuberculosis during infection, being required for virulence after entering cells via the amino acid transporter AnsP1.
Cytoplasmic polyadenylation activates quiescent transcripts for translation by selective poly(A)-tail extension. An approach involving a clickable adenosine derivative permits capture of newly polyadenylated transcripts, and next-generation sequencing reveals mRNA sequence motifs that are linked to polyadenylation.
In cells, di-iron hydrogenases require three maturases to facilitate proper assembly of metal clusters. Reconstitution experiments with synthetic cofactor mimics coupled with functional and spectroscopic characterization now show these helper proteins are not needed in vitro to form highly active H2-producing catalysts.
Fosfomycin inhibits cell wall formation by preventing MurA-mediated UDP-MurNAc synthesis, but resistance to this drug suggested another route to UDP-MurNAc might exist. Genetic and biochemical studies identify two genes that, with an unknown phosphatase, define a new MurNAc salvage pathway.
The exchange of a heme-ligating cysteine for a serine residue in an engineered P450 alters the redox potential of the enzyme, deactivating wild-type function and enabling efficient, NADPH-mediated carbene transfer in cells.
Carbenes have been postulated to take part in the catalytic cycle of several enzymes, but direct detection of these unstable compounds has been elusive. Spectroscopic and structural studies of pyruvate oxidase now identify a carbene-containing cofactor, calling for reinspection of existing enzyme mechanisms.
PreQ1, a metabolic precursor of the modified tRNA nucleotide queuosine, regulates expression of queuosine biosynthetic genes via two distinct classes of preQ1 riboswitch. Biochemical studies and the first X-ray crystal structure of a class II preQ1 riboswitch reveal how preQ1 recognizes this RNA motif and how it may regulate translation of downstream genes.
Analysis of proteins within their native environment can confirm and extend in vitro–derived conclusions. NMR analysis of superoxide dismutase 1 in live human cells now corroborates previously identified steps on the maturation pathway and demonstrates copper-independent function of the chaperone CCS.
Hedgehog acyltransferase (Hhat) attaches a palmitate to Sonic hedgehog, but the importance of this enzyme has been difficult to discern. RU-SKI 43 is a potent and selective inhibitor of Hhat in vitro and in cells that will allow scientists to investigate the importance of Hhat for Shh signaling.
The VAS1 aminotransferase regulates the shade avoidance response in Arabidopsis thaliana by shunting metabolic flux away from auxin and ethylene, two growth regulatory plant hormones.
Bacteria find creative solutions to occupy a variety of environmental niches. A peptide isolated from a gold-associated microbe provides a new example of this adaptation, binding gold and precipitating it to protect the organism from metal toxicity.
Heterologous expression of the two components of enterococcal cytolysin with a lanthionine synthetase enables the structural determination of this antimicrobial and hemolytic pair, revealing unexpected stereochemistry in one of the lanthionine bridges that is driven by peptide sequence.
A natural aliphatic alcohol ligand of the mouse orphan odorant receptor Olfr288 from the male preputial gland is regulated by testosterone and affects attractiveness to female mice.
Ubiquitin-conjugating (E2) enzymes contain a conserved asparagine that has been proposed to stabilize an oxyanion intermediate. Structural and biochemical studies of Ubc13 suggest that this residue has a structural role in stabilizing the E2 active site.
