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| Status |
Public on Mar 24, 2009 |
| Title |
E2-RING expansion of the NEDD8 cascade confers specificity to cullin modification |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Ubiquitin and ubiquitin-like proteins (UBLs) are directed to targets by cascades of E1, E2, and E3 enzymes. The largest ubiquitin E3 subclass consists of cullin-RING ligases (CRLs), which contain one each of several cullins (CUL1, -2, -3, -4, or -5) and RING proteins (RBX1 or -2). CRLs are activated by ligation of the UBL NEDD8 to a conserved cullin lysine. How is cullin NEDD8ylation specificity established? Here we report that, like UBE2M (also known as UBC12), the previously uncharacterized E2 UBE2F is a NEDD8-conjugating enzyme in vitro and in vivo. Biochemical and structural analyses indicate how plasticity of hydrophobic E1-E2 interactions and E1 conformational flexibility allow one E1 to charge multiple E2s. The E2s have distinct functions, with UBE2M/RBX1 and UBE2F/RBX2 displaying different target cullin specificities. Together, these studies reveal the molecular basis for and functional importance of hierarchical expansion of the NEDD8 conjugation system in establishing selective CRL activation.
Mol Cell 33:483-495, 2009.
Keywords: Comparison of gene expression profiles
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| Overall design |
Ube2m and Ube2f are E2 enzymes that direct protein modification by NEDD8. Here we explore the specific functions of Ube2f and Ube2m by comparing gene expression profiles following knockdown of their function in NIH 3T3 cells.
We used microarrays to detail the global programme of gene expression changes following knockdown of Ube2m and Ube2f in NIH 3T3 cells.
NIH 3T3 cells were transduced with retroviral constructs containing shRNA directed against Ube2m or Ube2f. Three replicates of each condition were analyzed.
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| Contributor(s) |
Ayrault O, Huang DT, Neale GA, Murray PJ, Roussel MF, Schulman BA |
| Citation(s) |
19250909 |
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| Submission date |
Dec 22, 2008 |
| Last update date |
Feb 11, 2019 |
| Contact name |
Geoffrey Neale |
| E-mail(s) |
geoffrey.neale@stjude.org
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| Organization name |
St Jude Childrens Research Hospital
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| Department |
Hartwell Center
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| Street address |
262 Danny Thomas Place
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| City |
Memphis |
| State/province |
TN |
| ZIP/Postal code |
38105 |
| Country |
USA |
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| Platforms (1) |
| GPL1261 |
[Mouse430_2] Affymetrix Mouse Genome 430 2.0 Array |
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| Samples (9)
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| GSM353503 |
control shRNA, biological replicate 1 |
| GSM353504 |
control shRNA, biological replicate 2 |
| GSM353505 |
Ube2m shRNA, biological replicate 1 |
| GSM353506 |
Ube2m shRNA, biological replicate 2 |
| GSM353507 |
Ube2f shRNA, biological replicate 1 |
| GSM353508 |
Ube2f shRNA, biological replicate 2 |
| GSM353509 |
control shRNA, biological replicate 3 |
| GSM353510 |
Ube2m shRNA, biological replicate 3 |
| GSM353511 |
Ube2f shRNA, biological replicate 3 |
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| Relations |
| BioProject |
PRJNA112515 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE14088_RAW.tar |
35.5 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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