title: Regulation of E2F1 by APC/C Cdh1 via K11 linkage-specific ubiquitin chain formation.
abstract: E2F1 is a eukaryotic transcription factor that is known to regulate various cellular pathways such as cell cycle progression, DNA replication, DNA damage responses and induction of apoptosis. Given its versatile roles, a precise and tight regulation of E2F1 is very critical to maintain genomic stability. E2F1 is regulated both at transcriptional and posttranslational levels during cell cycle and upon DNA damage. After S phase, E2F1 is targeted for degradation and is kept at low levels or in an inactive state until the next G 1/S phase transition. Our studies show that APC/C ubiquitin ligase in conjunction with its co-activator Cdh1 (APC/C (Cdh1) ) can downregulate E2F1. We also identify an APC/C subunit APC5 that binds to E2F1 and is essential for E2F1 ubiquitination. We confirm an interaction between E2F1 and Cdh1 as well as an interaction between E2F1 and APC5 both in vivo and in vitro. In vitro GST pull-down assays have mapped the C-terminal 79 a.a. of E2F1 as Cdh1 interacting residues. Ectopically expressed Cdh1 downregulates the expression of E2F1-4. Our studies have also shown for the first time that E2F1 can be modified by K11-linkage specific ubiquitin chain formation (Ub-K11). The formation of Ub-K11 chains on E2F1 is increased in the presence of Cdh1 and accumulated in the presence of proteasome inhibitor, suggesting that APC/C (Cdh1)  targets E2F1 for degradation by forming Ub-K11 chains. We also show that the effect of Cdh1 on E2F1 degradation is blocked upon DNA damage. Interestingly, Ub-K11-linked E2F1 accumulates after treatment of DNA damaging agents. The data suggest that DNA damage signaling processes do not inhibit APC/C (Cdh1)  to ubiquitinate E2F1. Instead, they block the proteasomal degradation of Ub-K11-linked E2F1, and therefore lead to its accumulation.
authors.affiliation: Section of Hematology/Oncology, Department of Medicine, Baylor College of Medicine, Houston, TX, USA.
authors.forename: Varija N
authors.initials: VN
authors.lastname: Budhavarapu
authors.forename: Erin D
authors.initials: ED
authors.lastname: White
authors.forename: Christina S
authors.initials: CS
authors.lastname: Mahanic
authors.forename: Ligong
authors.initials: L
authors.lastname: Chen
authors.forename: Fang-Tsyr
authors.initials: FT
authors.lastname: Lin
authors.forename: Weei-Chin
authors.initials: WC
authors.lastname: Lin
mesh_terms: D064173:Anaphase-Promoting Complex-Cyclosome; D015703:Antigens, CD; D064193:Apc5 Subunit, Anaphase-Promoting Complex-Cyclosome; D015820:Cadherins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D004249:DNA Damage; D015536:Down-Regulation; D050687:E2F1 Transcription Factor / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D011485:Protein Binding; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D016196:S Phase; D025801:Ubiquitin / Q000737:chemistry / Q000378:metabolism*; D043743:Ubiquitin-Protein Ligase Complexes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination
chemical_list: C577375:ANAPC5 protein, human; D015703:Antigens, CD; D064193:Apc5 Subunit, Anaphase-Promoting Complex-Cyclosome; C513384:CDH1 protein, human; D015820:Cadherins; D050687:E2F1 Transcription Factor; D021122:Protein Subunits; D034741:RNA, Small Interfering; D025801:Ubiquitin; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome