Spindle Assembly Checkpoint Protein Cdc20 Transcriptionally Activates Expression of Ubiquitin Carrier Protein UbcH10

Nath, Somsubhra; Banerjee, Taraswi; Sen, Debrup; Das, Tania; Roychoudhury, Susanta

doi:10.1074/jbc.m110.160671

Cited by 26 publications

(32 citation statements)

References 54 publications

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“…Plasmids and siRNAs Used-Different deletion constructs of the UBE2C promoter containing the Ϫ693/ϩ39, Ϫ321/ϩ39, or Ϫ141/ϩ39-bp fragments across the transcription start site were prepared as described previously (34). The site-directed mutations of the CCAAT and CHR elements on the Ϫ141/ ϩ39-bp UBE2C promoter were created using the site-directed mutagenesis kit (Stratagene, La Jolla, CA).…”

Section: Methodsmentioning

confidence: 99%

E2 Ubiquitin-conjugating Enzyme, UBE2C Gene, Is Reciprocally Regulated by Wild-type and Gain-of-Function Mutant p53

Bajaj

Alam

Roy

et al. 2016

Journal of Biological Chemistry

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Spindle assembly checkpoint governs proper chromosomal segregation during mitosis to ensure genomic stability. At the cellular level, this event is tightly regulated by UBE2C, an E2 ubiquitin-conjugating enzyme that donates ubiquitin to the anaphase-promoting complex/cyclosome. This, in turn, facilitates anaphase-onset by ubiquitin-mediated degradation of mitotic substrates. UBE2C is an important marker of chromosomal instability and has been associated with malignant growth. However, the mechanism of its regulation is largely unexplored. In this study, we report that UBE2C is transcriptionally activated by the gain-of-function (GOF) mutant p53, although it is transcriptionally repressed by wild-type p53. We showed that wild-type p53-mediated inhibition of UBE2C is p21-E2F4-dependent and GOF mutant p53-mediated transactivation of UBE2C is NF-Y-dependent. We further explored that DNA damage-induced wild-type p53 leads to spindle assembly checkpoint arrest by repressing UBE2C, whereas mutant p53 causes premature anaphase exit by increasing UBE2C expression in the presence of 5-fluorouracil. Identification of UBE2C as a target of wild-type and GOF mutant p53 further highlights the contribution of p53 in regulation of spindle assembly checkpoint.

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Section: Methodsmentioning

confidence: 99%

E2 Ubiquitin-conjugating Enzyme, UBE2C Gene, Is Reciprocally Regulated by Wild-type and Gain-of-Function Mutant p53

Bajaj

Alam

Roy

et al. 2016

Journal of Biological Chemistry

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“…Different UBCH10 promoter-luciferase gene constructs (pSN1 to pSN4) were described previously (35). The E2F1 binding site deletion mutant (pSN5) was created from the pSN2 construct by use of a QuikChange XL site-directed mutagenesis kit (Stratagene, La Jolla, CA).…”

Section: Methodsmentioning

confidence: 99%

“…Quantitative real-time PCR was done as described previously (35). Primer sets for E2F1, UBCH10, and GAPDH are listed in Table S4 in the supplemental material.…”

Section: Methodsmentioning

confidence: 99%

“…We hypothesized that overexpressed Cdc20 requires upregulated UbcH10 expression, which in turn prematurely ubiquitinates and activates Cdc20, resulting in the abnormal mitotic progression observed in cancer cells. Interestingly, we found that Cdc20 itself acts as a transcriptional coactivator and upregulates UbcH10 in a cell cycle-specific manner (35). The previous study showed that Cdc20-APC/C along with the transcriptional coactivator CBP/p300 binds to the response element (RE) sequence in the UBCH10 promoter and upregulates its transcription.…”

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confidence: 99%

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Deregulation of Rb-E2F1 Axis Causes Chromosomal Instability by Engaging the Transactivation Function of Cdc20–Anaphase-Promoting Complex/Cyclosome

Nath

Chowdhury

Dey

et al. 2015

Molecular and Cellular Biology

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The E2F family of transcription factors regulates genes involved in various aspects of the cell cycle. Beyond the well-documented role in G 1 /S transition, mitotic regulation by E2F has also been reported. Proper mitotic progression is monitored by the spindle assembly checkpoint (SAC). The SAC ensures bipolar separation of chromosomes and thus prevents aneuploidy. There are limited reports on the regulation of the SAC by E2F. Our previous work identified the SAC protein Cdc20 as a novel transcriptional regulator of the mitotic ubiquitin carrier protein UbcH10. However, none of the Cdc20 transcription complex proteins have any known DNA binding domain. Here we show that an E2F1-DP1 heterodimer is involved in recruitment of the Cdc20 transcription complex to the UBCH10 promoter and in transactivation of the gene. We further show that inactivation of Rb can facilitate this transactivation process. Moreover, this E2F1-mediated regulation of UbcH10 influences mitotic progression. Deregulation of this pathway results in premature anaphase, chromosomal abnormalities, and aneuploidy. We conclude that excess E2F1 due to Rb inactivation recruits the complex of Cdc20 and the anaphase-promoting complex/cyclosome (Cdc20-APC/C) to deregulate the expression of UBCH10, leading to chromosomal instability in cancer cells.

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“…It is noteworthy that Cdc20 levels impact both mitotic slippage rate and cell fate. 29,31,32 Meanwhile, inhibiting endogenous miR-125b resulted in faster mitotic exit, possibly by inactivating SAC through increased Mad1 levels. This is reminiscent of a recent report that Mad1 upregulation might promote tumour miR-125b represses MAD1 expression S Bhattacharjya et al formation and manifest resistance to therapies.…”

Section: Discussionmentioning

confidence: 99%

miR-125b promotes cell death by targeting spindle assembly checkpoint gene MAD1 and modulating mitotic progression

et al. 2012

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The spindle assembly checkpoint (SAC) is a 'wait-anaphase' mechanism that has evolved in eukaryotic cells in response to the stochastic nature of chromosome-spindle attachments. In the recent past, different aspects of the SAC regulation have been described. However, the role of microRNAs in the SAC is vaguely understood. We report here that Mad1, a core SAC protein, is repressed by human miR-125b. Mad1 serves as an adaptor protein for Mad2 -which functions to inhibit anaphase entry till the chromosomal defects in metaphase are corrected. We show that exogenous expression of miR-125b, through downregulation of Mad1, delays cells at metaphase. As a result of this delay, cells proceed towards apoptotic death, which follows from elevated chromosomal abnormalities upon ectopic expression of miR-125b. Moreover, expressions of Mad1 and miR-125b are inversely correlated in a variety of cancer cell lines, as well as in primary head and neck tumour tissues. We conclude that increased expression of miR-125b inhibits cell proliferation by suppressing Mad1 and activating the SAC transiently. We hypothesize an optimum Mad1 level and thus, a properly scheduled SAC is maintained partly by miR-125b.

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Spindle Assembly Checkpoint Protein Cdc20 Transcriptionally Activates Expression of Ubiquitin Carrier Protein UbcH10

Cited by 26 publications

References 54 publications

E2 Ubiquitin-conjugating Enzyme, UBE2C Gene, Is Reciprocally Regulated by Wild-type and Gain-of-Function Mutant p53

E2 Ubiquitin-conjugating Enzyme, UBE2C Gene, Is Reciprocally Regulated by Wild-type and Gain-of-Function Mutant p53

Deregulation of Rb-E2F1 Axis Causes Chromosomal Instability by Engaging the Transactivation Function of Cdc20–Anaphase-Promoting Complex/Cyclosome

miR-125b promotes cell death by targeting spindle assembly checkpoint gene MAD1 and modulating mitotic progression

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