title: Nuclear α Spectrin Differentially Affects Monoubiquitinated Versus Non-Ubiquitinated FANCD2 Function After DNA Interstrand Cross-Link Damage.
abstract: Nonerythroid α spectrin (αIISp) and the Fanconi anemia (FA) protein, FANCD2, play critical roles in DNA interstrand cross-link (ICL) repair during S phase. Both are needed for recruitment of repair proteins, such as XPF, to sites of damage and repair of ICLs. However, the relationship between them in ICL repair and whether αIISp is involved in FANCD2's function in repair is unclear. The present studies show that, after ICL formation, FANCD2 disassociates from αIISp and localizes, before αIISp, at sites of damage in nuclear foci. αIISp and FANCD2 foci do not co-localize, in contrast to our previous finding that αIISp and the ICL repair protein, XPF, co-localize and follow a similar time course for formation. Knock-down of αIISp has no effect on monoubiquitination of FANCD2 (FANCD2-Ub) or its localization to chromatin or foci, though it leads to decreased ICL repair. Studies using cells from FA patients, defective in ICL repair and αIISp, have elucidated an important role for αIISp in the function of non-Ub FANCD2. In FA complementation group A (FA-A) cells, in which FANCD2 is not monoubiquitinated and does not form damage-induced foci, we demonstrate that restoration of αIISp levels to normal, by knocking down the protease μ-calpain, leads to formation of non-Ub FANCD2 foci after ICL damage. Since restoration of αIISp levels in FA-A cells restores DNA repair and cell survival, we propose that αIISp is critical for recruitment of non-Ub FANCD2 to sites of damage, which has an important role in the repair response and ICL repair.
authors: Department of Pathology and Laboratory Medicine, Rutgers New Jersey Medical School, 185 South Orange Avenue, Newark, New Jersey, 07103, USA.
authors: Pan
authors: P
authors: Zhang
authors: Department of Pathology and Laboratory Medicine, Rutgers New Jersey Medical School, 185 South Orange Avenue, Newark, New Jersey, 07103, USA.
authors: Deepa
authors: D
authors: Sridharan
authors: Department of Pathology and Laboratory Medicine, Rutgers New Jersey Medical School, 185 South Orange Avenue, Newark, New Jersey, 07103, USA.
authors: Muriel W
authors: MW
authors: Lambert
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000378:metabolism*; D006801:Humans; D021381:Protein Transport; D013049:Spectrin / Q000378:metabolism*; D057149:Ubiquitinated Proteins / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: D002843:Chromatin; D004268:DNA-Binding Proteins; C499905:FANCD2 protein, human; D052236:Fanconi Anemia Complementation Group D2 Protein; D057149:Ubiquitinated Proteins; C095888:xeroderma pigmentosum group F protein; D013049:Spectrin
keywords: DNA DAMAGE; DNA INTERSTRAND CROSS-LINKS; DNA REPAIR; FANCD2; FANCONI ANEMIA; NON-UBIQUITINATED-FANCD2; NONERYTHROID ALPHA SPECTRIN; NUCLEAR SPECTRIN; XPF