TY - JOUR
T1 - Solution structure and dynamics of yeast elongin C in complex with a von Hippel-Lindau peptide
AU - Botuyan, Maria Victoria
AU - Mer, Georges
AU - Yi, Gwan Su
AU - Koth, Christopher M.
AU - Case, David A.
AU - Edwards, Aled M.
AU - Chazin, Walter J.
AU - Arrowsmith, Cheryl H.
N1 - Funding Information:
We thank John Chung for help in the setup of NMR experiments; Adelinda Yee, Gerard Kroon, Micah Gearhart, Brendan Duggan and Signe Holmbeck for assistance with NMRPipe/NMRView; and Gerard Kroon for assistance with Modelfree. This research was supported by grants from the National Cancer Institute of Canada to C.H.A. and A.M.E, the National Science Foundation and Vanderbilt University Molecular Toxicology and Ingram Cancer Centers to W.J.C., and the National Institutes of Health (GM45811) to D.A.C.
PY - 2001/9/7
Y1 - 2001/9/7
N2 - Elongin is a transcription elongation factor that stimulates the rate of elongation by suppressing transient pausing by RNA polymerase II at many sites along the DNA. It is heterotrimeric in mammals, consisting of elongins A, B and C subunits, and bears overall similarity to a class of E3 ubiquitin ligases known as SCF (Skp1-Cdc53 (cullin)-F-box) complexes. A subcomplex of elongins B and C is a target for negative regulation by the von Hippel-Lindau (VHL) tumor-suppressor protein. Elongin C from Saccharomyces cerevisiae, Elc1, exhibits high sequence similarity to mammalian elongin C. Using NMR spectroscopy we have determined the three-dimensional structure of Elc1 in complex with a human VHL peptide, VHL(157-171), representing the major Elc1 binding site. The bound VHL peptide is entirely helical. Elc1 utilizes two C-terminal helices and an intervening loop to form a binding groove that fits VHL(157-171). Chemical shift perturbation and dynamics analyses reveal that a global conformational change accompanies Elc1/VHL(157-171) complex formation. Moreover, the disappearance of conformational exchange phenomena on the microsecond to millisecond time scale within Eld upon VHL peptide binding suggests a role for slow internal motions in ligand recognition.
AB - Elongin is a transcription elongation factor that stimulates the rate of elongation by suppressing transient pausing by RNA polymerase II at many sites along the DNA. It is heterotrimeric in mammals, consisting of elongins A, B and C subunits, and bears overall similarity to a class of E3 ubiquitin ligases known as SCF (Skp1-Cdc53 (cullin)-F-box) complexes. A subcomplex of elongins B and C is a target for negative regulation by the von Hippel-Lindau (VHL) tumor-suppressor protein. Elongin C from Saccharomyces cerevisiae, Elc1, exhibits high sequence similarity to mammalian elongin C. Using NMR spectroscopy we have determined the three-dimensional structure of Elc1 in complex with a human VHL peptide, VHL(157-171), representing the major Elc1 binding site. The bound VHL peptide is entirely helical. Elc1 utilizes two C-terminal helices and an intervening loop to form a binding groove that fits VHL(157-171). Chemical shift perturbation and dynamics analyses reveal that a global conformational change accompanies Elc1/VHL(157-171) complex formation. Moreover, the disappearance of conformational exchange phenomena on the microsecond to millisecond time scale within Eld upon VHL peptide binding suggests a role for slow internal motions in ligand recognition.
KW - Conformational change
KW - Elongin
KW - Ligand recognition
KW - NMR solution structure
KW - Von Hippel-Lindau
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U2 - 10.1006/jmbi.2001.4938
DO - 10.1006/jmbi.2001.4938
M3 - Article
C2 - 11545595
AN - SCOPUS:0035823058
SN - 0022-2836
VL - 312
SP - 177
EP - 186
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -