Tangled up in knots: structures of inactivated forms of E. coli class Ia ribonucleotide reductase.
Ribonucleotide reductases (RNRs) provide the precursors for DNA biosynthesis and repair and are successful targets for anticancer drugs such as clofarabine and gemcitabine. Recently, we reported that dATP inhibits E. coli class Ia RNR by driving formation of RNR subunits into α4β4 rings. Here, we present the first X-ray structure of a gemcitabine-inhibited E. coli RNR and show that the previously described α4β4 rings can interlock to form an unprecedented (α4β4)2 megacomplex. This complex is also seen in a higher-resolution dATP-inhibited RNR structure presented here, which employs a distinct crystal lattice from that observed in the gemcitabine-inhibited case. With few reported examples of protein catenanes, we use data from small-angle X-ray scattering and electron microscopy to both understand the solution conditions that contribute to concatenation in RNRs as well as present a mechanism for the formation of these unusual structures.
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Christina M
CM
Zimanyi
Nozomi
N
Ando
Edward J
EJ
Brignole
Francisco J
FJ
Asturias
Joanne
J
Stubbe
Catherine L
CL
Drennan
D018360:Crystallography, X-Ray; D003565:Cytidine Diphosphate / Q000031:analogs & derivatives / Q000737:chemistry; D003838:Deoxyadenine Nucleotides / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000648:ultrastructure; D005095:Exoribonucleases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000737:chemistry; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction
D003838:Deoxyadenine Nucleotides; D029968:Escherichia coli Proteins; D021122:Protein Subunits; C523676:gemcitabine-5'-diphosphate; C489683:rnr protein, E coli; D003565:Cytidine Diphosphate; D005095:Exoribonucleases; C026600:2'-deoxyadenosine triphosphate