Downregulation of the Cytosolic Iron-Sulfur Assembly Pathway in Cancer by an E3 Ubiquitin Ligase
| dc.contributor.advisor | Tu, Benjamin | en |
| dc.contributor.committeeMember | Potts, Patrick Ryan | en |
| dc.contributor.committeeMember | Minna, John D. | en |
| dc.contributor.committeeMember | Liu, Yi | en |
| dc.creator | Weon, Jenny Linda | en |
| dc.creator.orcid | 0000-0002-4815-6659 | |
| dc.date.accessioned | 2021-06-03T22:29:43Z | |
| dc.date.available | 2021-06-03T22:29:43Z | |
| dc.date.created | 2019-05 | |
| dc.date.issued | 2017-06-19 | |
| dc.date.submitted | May 2019 | |
| dc.date.updated | 2021-06-03T22:29:43Z | |
| dc.description.abstract | Iron-sulfur (Fe-S) clusters are considered to be one of the oldest cofactors utilized by proteins and are essential for life from bacteria to mammals. Multiple processes in the cell require Fe-S cofactors, such as electron transfer in mitochondrial respiration, enzymatic reactions, and as structural components in DNA repair enzymes. We describe here the first post-translational mechanism to regulate Fe-S assembly and delivery through the ubiquitination and degradation of a key cytosolic iron-sulfur cluster assembly (CIA) pathway component by a MAGE-RING ligase (MRL). The MAGE protein family consists of ~40 members in humans that function in complex with E3 ubiquitin ligases to enhance ubiquitination activity, alter E3 subcellular localization, and/or specify E3 targets. Using biochemical and cellular approaches we have discovered that the MAGE-F1-NSE1 ligase disrupts Fe-S cluster delivery through ubiquitination and degradation of the CIA pathway protein MMS19. MMS19 is a substrate specifying, late-acting component of the CIA pathway that facilitates Fe-S transfer from the multi-component cascade of assembly proteins to specific recipient apoproteins. Notably, many MMS19 targets are enzymes involved in DNA repair. We found that MAGE-F1 directs the E3 ligase NSE1 to target MMS19 for ubiquitination and degradation. Knockdown of MAGE-F1 stabilized MMS19 and overexpression of MAGE-F1 decreased MMS19 levels without affecting MMS19 mRNA levels. We further confirmed MAGE-F1 inhibits Fe-S incorporation into known MMS19-dependent Fe-S proteins, such as FANCJ, POLD1, RTEL1, XPD, and DPYD, but not MMS19-independent Fe-S proteins, such as PPAT. Loss of Fe-S incorporation leads to decreased DNA repair capacity of cells, exemplified by decreased homologous recombination rates and altered sensitivity to DNA damaging agents. Importantly, numerous cancer types harbor copy-number amplification of MAGE-F1, including lung squamous carcinoma and head and neck squamous carcinoma. Consistent with MAGE-F1 inhibitory activity on Fe-S incorporation into key DNA repair enzymes, MAGE-F1-amplified tumors bear a significantly greater mutational burden than non-MAGE-F1-amplified cancers and the expression of MAGE-F1-NSE1 correlates with poor patient prognosis. In summary, we provide the first evidence for post-translational regulatory control of Fe-S cluster assembly and a novel mechanism by which a broad spectrum of DNA repair enzymes can be regulated and lead to genomic instability in cancer. | en |
| dc.format.mimetype | application/pdf | en |
| dc.identifier.oclc | 1255189290 | |
| dc.identifier.uri | https://hdl.handle.net/2152.5/9527 | |
| dc.language.iso | en | en |
| dc.subject | Carrier Proteins | en |
| dc.subject | DNA Repair | en |
| dc.subject | Iron | en |
| dc.subject | Microtubule-Associated Proteins | en |
| dc.subject | Neoplasm Proteins | en |
| dc.subject | Sulfur | en |
| dc.subject | Transcription Factors | en |
| dc.title | Downregulation of the Cytosolic Iron-Sulfur Assembly Pathway in Cancer by an E3 Ubiquitin Ligase | en |
| dc.type | Thesis | en |
| dc.type.material | text | en |
| thesis.degree.department | Graduate School of Biomedical Sciences | en |
| thesis.degree.discipline | Integrative Biology | en |
| thesis.degree.grantor | UT Southwestern Medical Center | en |
| thesis.degree.level | Doctoral | en |
| thesis.degree.name | Doctor of Philosophy | en |