title: Dynamics and metal exchange properties of C4C4 RING domains from CNOT4 and the p44 subunit of TFIIH.
abstract: Zinc fingers are small structured protein domains that require the coordination of zinc for a stable tertiary fold. Together with FYVE and PHD, the RING domain forms a distinct class of zinc-binding domains, where two zinc ions are ligated in a cross-braced manner, with the first and third pairs of ligands coordinating one zinc ion, while the second and fourth pairs ligate the other zinc ion. To investigate the relationship between the stability and dynamic behaviour of the domains and the stability of the metal-binding site, we studied metal exchange for the C4C4 RING domains of CNOT4 and the p44 subunit of TFIIH. We found that Zn(2+)-Cd(2+) exchange is different between the two metal-binding sites in the C4C4 RING domains of the two proteins. In order to understand the origins of these distinct exchange rates, we studied the backbone dynamics of both domains in the presence of zinc and of cadmium by NMR spectroscopy. The differential stability of the two metal-binding sites in the RING domains, as reflected by the different metal exchange rates, can be explained by a combination of accessibility and an electrostatic ion interaction model. A greater backbone flexibility for the p44 RING domain as compared to CNOT4 may be related to the distinct types of protein-protein interactions in which the two C4C4 RING domains are involved.
authors.affiliation: Department of NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
authors.forename: Klaartje
authors.initials: K
authors.lastname: Houben
authors.forename: Emeric
authors.initials: E
authors.lastname: Wasielewski
authors.forename: Cyril
authors.initials: C
authors.lastname: Dominguez
authors.forename: Esther
authors.initials: E
authors.lastname: Kellenberger
authors.forename: R Andrew
authors.initials: RA
authors.lastname: Atkinson
authors.forename: H Th Marc
authors.initials: HT
authors.lastname: Timmers
authors.forename: Bruno
authors.initials: B
authors.lastname: Kieffer
authors.forename: Rolf
authors.initials: R
authors.lastname: Boelens
mesh_terms: D000595:Amino Acid Sequence; D002104:Cadmium / Q000378:metabolism; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D017434:Protein Structure, Tertiary; D051758:Transcription Factor TFIIH; D014157:Transcription Factors; D019473:Transcription Factors, TFII / Q000737:chemistry / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism
chemical_list: C501331:CNOT4 protein, human; D009587:Nitrogen Isotopes; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D002104:Cadmium; D051758:Transcription Factor TFIIH; D015032:Zinc