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Science 23 April 1999:
Vol. 284. no. 5414, pp. 662 - 665
DOI: 10.1126/science.284.5414.662
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Reports

Reconstitution of G1 Cyclin Ubiquitination with Complexes Containing SCFGrr1 and Rbx1

Dorota Skowyra, 1 Deanna M. Koepp, 123 Takumi Kamura, 45 Michael N. Conrad, 5 Ronald C. Conaway, 5 Joan Weliky Conaway, 456 Stephen J. Elledge, 123 J. Wade Harper 1*

Control of cyclin levels is critical for proper cell cycle regulation. In yeast, the stability of the G1 cyclin Cln1 is controlled by phosphorylation-dependent ubiquitination. Here it is shown that this reaction can be reconstituted in vitro with an SCF E3 ubiquitin ligase complex. Phosphorylated Cln1 was ubiquitinated by SCF (Skp1-Cdc53-F-box protein) complexes containing the F-box protein Grr1, Rbx1, and the E2 Cdc34. Rbx1 promotes association of Cdc34 with Cdc53 and stimulates Cdc34 auto-ubiquitination in the context of Cdc53 or SCF complexes. Rbx1, which is also a component of the von Hippel-Lindau tumor suppressor complex, may define a previously unrecognized class of E3-associated proteins.

1 Verna and Marrs McLean Department of Biochemistry,
2 Department of Molecular and Human Genetics,
3 Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030, USA.
4 Howard Hughes Medical Institute and
5 Program in Molecular and Cell Biology, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA.
6 Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73190, USA.
*   To whom correspondence should be addressed. E-mail: jharper{at}bcm.tmc.edu.

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F-Box Protein Grr1 Interacts with Phosphorylated Targets via the Cationic Surface of Its Leucine-Rich Repeat.
Y. G. Hsiung, H.-C. Chang, J.-L. Pellequer, R. La Valle, S. Lanker, and C. Wittenberg (2001)
Mol. Cell. Biol. 21, 2506-2520
   Abstract »    Full Text »
ATF4 Degradation Relies on a Phosphorylation-Dependent Interaction with the SCF{beta}TrCP Ubiquitin Ligase.
I. Lassot, E. Ségéral, C. Berlioz-Torrent, H. Durand, L. Groussin, T. Hai, R. Benarous, and F. Margottin-Goguet (2001)
Mol. Cell. Biol. 21, 2192-2202
   Abstract »    Full Text »
Interaction of the E1A Oncoprotein with Yak1p, a Novel Regulator of Yeast Pseudohyphal Differentiation, and Related Mammalian Kinases.
Z. Zhang, M. M. Smith, and J. S. Mymryk (2001)
Mol. Biol. Cell 12, 699-710
   Abstract »    Full Text »
Phosphorylation- and SKP1-independent in Vitro Ubiquitination of E2F1 by Multiple ROC-Cullin Ligases.
T. Ohta and Y. Xiong (2001)
Cancer Res. 61, 1347-1353
   Abstract »    Full Text »
Rma1, a novel type of RING finger protein conserved from Arabidopsis to human, is a membrane-bound ubiquitin ligase.
N Matsuda, T Suzuki, K Tanaka, and A Nakano (2001)
J. Cell Sci. 114, 1949-1957
   Abstract »    PDF »
The Cell Biology of the COP/DET/FUS Proteins. Regulating Proteolysis in Photomorphogenesis and Beyond?.
C. S. Hardtke and X.-W. Deng (2000)
Plant Physiology 124, 1548-1557
   Full Text »
The Abundance of Met30p Limits SCFMet30p Complex Activity and Is Regulated by Methionine Availability.
D. B. Smothers, L. Kozubowski, C. Dixon, M. G. Goebl, and N. Mathias (2000)
Mol. Cell. Biol. 20, 7845-7852
   Abstract »    Full Text »
The CUL1 C-Terminal Sequence and ROC1 Are Required for Efficient Nuclear Accumulation, NEDD8 Modification, and Ubiquitin Ligase Activity of CUL1.
M. Furukawa, Y. Zhang, J. McCarville, T. Ohta, and Y. Xiong (2000)
Mol. Cell. Biol. 20, 8185-8197
   Abstract »    Full Text »
Alphaherpesvirus Proteins Related to Herpes Simplex Virus Type 1 ICP0 Affect Cellular Structures and Proteins.
J. Parkinson and R. D. Everett (2000)
J. Virol. 74, 10006-10017
   Abstract »    Full Text »
Endoplasmic Reticulum Degradation Requires Lumen to Cytosol Signaling: Transmembrane Control of Hrd1p by Hrd3p.
R. G. Gardner, G. M. Swarbrick, N. W. Bays, S. R. Cronin, S. Wilhovsky, L. Seelig, C. Kim, and R. Y. Hampton (2000)
J. Cell Biol. 151, 69-82
   Abstract »    Full Text »    PDF »
Activation of HIF1alpha ubiquitination by a reconstituted von Hippel-Lindau (VHL) tumor suppressor complex.
T. Kamura, S. Sato, K. Iwai, M. Czyzyk-Krzeska, R. C. Conaway, and J. W. Conaway (2000)
PNAS
   Abstract »    Full Text »
The RING-H2 finger protein APC11 and the E2 enzyme UBC4 are sufficient to ubiquitinate substrates of the anaphase-promoting complex.
M. Gmachl, C. Gieffers, A. V. Podtelejnikov, M. Mann, and J.-M. Peters (2000)
PNAS 97, 8973-8978
   Abstract »    Full Text »    PDF »
Cell Cycle-Dependent Expression of Mammalian E2-C Regulated by the Anaphase-Promoting Complex/Cyclosome.
A. Yamanaka, S. Hatakeyama, K.-i. Kominami, M. Kitagawa, M. Matsumoto, and K.-i. Nakayama (2000)
Mol. Biol. Cell 11, 2821-2831
   Abstract »    Full Text »
Human Papillomavirus Type 16 E6 Induces Self-Ubiquitination of the E6AP Ubiquitin-Protein Ligase.
W. H. Kao, S. L. Beaudenon, A. L. Talis, J. M. Huibregtse, and P. M. Howley (2000)
J. Virol. 74, 6408-6417
   Abstract »    Full Text »
Hsl1p, a Swe1p Inhibitor, Is Degraded via the Anaphase-Promoting Complex.
J. L. Burton and M. J. Solomon (2000)
Mol. Cell. Biol. 20, 4614-4625
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Isolation and Characterization of HRT1 Using a Genetic Screen for Mutants Unable to Degrade Gic2p in Saccharomyces cerevisiae.
M. Blondel, J.-M. Galan, and M. Peter (2000)
Genetics 155, 1033-1044
   Abstract »    Full Text »
Ariadne-1: A Vital Drosophila Gene Is Required in Development and Defines a New Conserved Family of RING-Finger Proteins.
M. Aguilera, M. Oliveros, M. Martínez-Padrón, J. A. Barbas, and A. Ferrús (2000)
Genetics 155, 1231-1244
   Abstract »    Full Text »
HRD Gene Dependence of Endoplasmic Reticulum-associated Degradation.
S. Wilhovsky, R. Gardner, and R. Hampton (2000)
Mol. Biol. Cell 11, 1697-1708
   Abstract »    Full Text »
Nedd8 Modification of Cul-1 Activates SCFbeta TrCP-Dependent Ubiquitination of Ikappa Balpha.
M. A. Read, J. E. Brownell, T. B. Gladysheva, M. Hottelet, L. A. Parent, M. B. Coggins, J. W. Pierce, V. N. Podust, R.-S. Luo, V. Chau, et al. (2000)
Mol. Cell. Biol. 20, 2326-2333
   Abstract »    Full Text »


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