UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary
Background & Aims: TP53 mutations underlie Barrett’s esophagus (BE) progression to dysplasia and cancer. During BE progression, the ubiquitin ligase (E3) RNF128/GRAIL switches expression from isoform 2 (Iso2) to Iso1, stabilizing mutant p53. However, the ubiquitin-conjugating enzyme (E2) tha...
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2022
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oai:doaj.org-article:4a56ad18c7804a3bbc16ac9b32c78f2e2021-11-14T04:34:11ZUBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary2352-345X10.1016/j.jcmgh.2021.08.003https://doaj.org/article/4a56ad18c7804a3bbc16ac9b32c78f2e2022-01-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2352345X21001703https://doaj.org/toc/2352-345XBackground & Aims: TP53 mutations underlie Barrett’s esophagus (BE) progression to dysplasia and cancer. During BE progression, the ubiquitin ligase (E3) RNF128/GRAIL switches expression from isoform 2 (Iso2) to Iso1, stabilizing mutant p53. However, the ubiquitin-conjugating enzyme (E2) that partners with Iso1 to stabilize mutant p53 is unknown. Methods: Single-cell RNA sequencing of paired normal esophagus and BE tissues identified candidate E2s, further investigated in expression data from BE to esophageal adenocarcinoma (EAC) progression samples. Biochemical and cellular studies helped clarify the role of RNF128-E2 on mutant p53 stability. Results: The UBE2D family member 2D3 (UBCH5C) is the most abundant E2 in normal esophagus. However, during BE to EAC progression, loss of UBE2D3 copy number and reduced expression of RNF128 Iso2 were noted, 2 known p53 degraders. In contrast, expression of UBE2D1 (UBCH5A) and RNF128 Iso1 in dysplastic BE and EAC forms an inactive E2–E3 complex, stabilizing mutant p53. To destabilize mutant p53, we targeted RNF128 Iso1 either by mutating asparagine (N48, 59, and 101) residues to block glycosylation to facilitate β-TrCP1–mediated degradation or by mutating proline (P54 and 105) residues to restore p53 polyubiquitinating ability. In addition, either loss of UBCH5A catalytic activity, or disruption of the Iso1-UBCH5A interaction promoted Iso1 loss. Consequently, overexpression of either catalytically dead or Iso1-binding–deficient UBCH5A mutants destabilized Iso1 to degrade mutant p53, thus compromising the clonogenic survival of mutant p53-dependent BE cells. Conclusions: Loss of RNF128 Iso2–UBCH5C and persistence of the Iso1–UBCH5A complex favors mutant p53 stability to promote BE cell survival. Therefore, targeting of Iso1-UBCH5A may provide a novel therapeutic strategy to prevent BE progression.Paramita RayDerek J. NancarrowDaysha Ferrer-TorresZhuwen WangMay San MartinhoTonaye HintonJoshua H. WuAngeline WuDanielle Kim TurgeonMax A. HammerMichael K. DameTheodore S. LawrencePatrick J. O’BrienJason R. SpenceDavid G. BeerDipankar RayElsevierarticleBarrett’s EsophagusEsophageal AdenocarcinomaRNF128-UBCH5 Complexp53 Protein StabilityDiseases of the digestive system. GastroenterologyRC799-869ENCellular and Molecular Gastroenterology and Hepatology, Vol 13, Iss 1, Pp 129-149 (2022) |
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EN |
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Barrett’s Esophagus Esophageal Adenocarcinoma RNF128-UBCH5 Complex p53 Protein Stability Diseases of the digestive system. Gastroenterology RC799-869 |
| spellingShingle |
Barrett’s Esophagus Esophageal Adenocarcinoma RNF128-UBCH5 Complex p53 Protein Stability Diseases of the digestive system. Gastroenterology RC799-869 Paramita Ray Derek J. Nancarrow Daysha Ferrer-Torres Zhuwen Wang May San Martinho Tonaye Hinton Joshua H. Wu Angeline Wu Danielle Kim Turgeon Max A. Hammer Michael K. Dame Theodore S. Lawrence Patrick J. O’Brien Jason R. Spence David G. Beer Dipankar Ray UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| description |
Background & Aims: TP53 mutations underlie Barrett’s esophagus (BE) progression to dysplasia and cancer. During BE progression, the ubiquitin ligase (E3) RNF128/GRAIL switches expression from isoform 2 (Iso2) to Iso1, stabilizing mutant p53. However, the ubiquitin-conjugating enzyme (E2) that partners with Iso1 to stabilize mutant p53 is unknown. Methods: Single-cell RNA sequencing of paired normal esophagus and BE tissues identified candidate E2s, further investigated in expression data from BE to esophageal adenocarcinoma (EAC) progression samples. Biochemical and cellular studies helped clarify the role of RNF128-E2 on mutant p53 stability. Results: The UBE2D family member 2D3 (UBCH5C) is the most abundant E2 in normal esophagus. However, during BE to EAC progression, loss of UBE2D3 copy number and reduced expression of RNF128 Iso2 were noted, 2 known p53 degraders. In contrast, expression of UBE2D1 (UBCH5A) and RNF128 Iso1 in dysplastic BE and EAC forms an inactive E2–E3 complex, stabilizing mutant p53. To destabilize mutant p53, we targeted RNF128 Iso1 either by mutating asparagine (N48, 59, and 101) residues to block glycosylation to facilitate β-TrCP1–mediated degradation or by mutating proline (P54 and 105) residues to restore p53 polyubiquitinating ability. In addition, either loss of UBCH5A catalytic activity, or disruption of the Iso1-UBCH5A interaction promoted Iso1 loss. Consequently, overexpression of either catalytically dead or Iso1-binding–deficient UBCH5A mutants destabilized Iso1 to degrade mutant p53, thus compromising the clonogenic survival of mutant p53-dependent BE cells. Conclusions: Loss of RNF128 Iso2–UBCH5C and persistence of the Iso1–UBCH5A complex favors mutant p53 stability to promote BE cell survival. Therefore, targeting of Iso1-UBCH5A may provide a novel therapeutic strategy to prevent BE progression. |
| format |
article |
| author |
Paramita Ray Derek J. Nancarrow Daysha Ferrer-Torres Zhuwen Wang May San Martinho Tonaye Hinton Joshua H. Wu Angeline Wu Danielle Kim Turgeon Max A. Hammer Michael K. Dame Theodore S. Lawrence Patrick J. O’Brien Jason R. Spence David G. Beer Dipankar Ray |
| author_facet |
Paramita Ray Derek J. Nancarrow Daysha Ferrer-Torres Zhuwen Wang May San Martinho Tonaye Hinton Joshua H. Wu Angeline Wu Danielle Kim Turgeon Max A. Hammer Michael K. Dame Theodore S. Lawrence Patrick J. O’Brien Jason R. Spence David G. Beer Dipankar Ray |
| author_sort |
Paramita Ray |
| title |
UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| title_short |
UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| title_full |
UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| title_fullStr |
UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| title_full_unstemmed |
UBCH5 Family Members Differentially Impact Stabilization of Mutant p53 via RNF128 Iso1 During Barrett’s Progression to Esophageal AdenocarcinomaSummary |
| title_sort |
ubch5 family members differentially impact stabilization of mutant p53 via rnf128 iso1 during barrett’s progression to esophageal adenocarcinomasummary |
| publisher |
Elsevier |
| publishDate |
2022 |
| url |
https://doaj.org/article/4a56ad18c7804a3bbc16ac9b32c78f2e |
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