ライフサイエンスコーパス: FANCI

1 FANCI and FANCD2 form the "ID" complex that loads onto c

2 FANCI is in close proximity to SF3B1 in the nucleoplasm

3 FANCI is integral to the Fanconi anemia (FA) pathway of

4 FANCI shares sequence similarity with FANCD2, likely evo

5 ANCD1 (1), FANCD2 (3), FANCF (2), FANCG (2), FANCI (1), FANCJ (2), and FANCL (3).

6 ions with RNA polymerase II on the BRCA1 and FANCI genes suggest a transcriptional defect in the abse

7 lized to viral replication compartments, and FANCI-D2 interacted with a multisubunit complex containi

8 FANCD2 and FANCI are thought to form a functional heterodimer durin

9 In chromatin, both FANCD2 and FANCI associate with SF3B1, prevent accumulation of post

10 followed by monoubiquitination of FANCD2 and FANCI by the FA core complex.

11 We propose that FANCD2 and FANCI contribute to the organization of functional domai

12 , monoubiquitination responses of FANCD2 and FANCI exhibit distinct DNA substrate specificities.

13 Our results suggest that FANCD2 and FANCI function separately at consecutive steps during DN

14 FANCD2 and FANCI function together in the Fanconi anemia network of

15 dimerization of monoubiquitinated FANCD2 and FANCI in chromatin is mediated in part through a noncova

16 ut cells display a unique lack of FANCD2 and FANCI localization to chromatin in exponentially growing

17 Monoubiquitinated FANCD2 and FANCI localize in chromatin-associated nuclear foci wher

18 he role of a proper DNA ligand in FANCD2 and FANCI monoubiquitination, and reveal regulatory mechanis

19 d PCNA stimulates FANCL-catalyzed FANCD2 and FANCI monoubiquitylation.

20 via the monoubiquitination of the FANCD2 and FANCI proteins, targeting these proteins to discrete nuc

21 ty of RAD18 in the recruitment of FANCD2 and FANCI to chromatin and the events leading to their ubiqu

22 ANCL, monoubiquitination of human FANCD2 and FANCI was examined.

23 s require Fanconi anemia proteins FANCD2 and FANCI, as well as Blm helicase, but not canonical DDR si

24 coni anemia (FA) pathway members, FANCD2 and FANCI, contribute to the repair of replication-stalling

25 rotein complex that ubiquitinates FANCD2 and FANCI, leading to formation of DNA repair structures.

26 lso play a role in mitosis, since FANCD2 and FANCI, the 2 key FA proteins, are localized to the extre

27 cruitment of two central players, FANCD2 and FANCI, to sites of stalled replication forks.

28 onsible for monoubiquitination of FANCD2 and FANCI.

29 ination of the downstream targets FANCD2 and FANCI.

30 alyzing the monoubiquitination of FANCD2 and FANCI.

31 red for the monoubiquitylation of FANCD2 and FANCI.

32 machine for monoubiquitination of FANCD2 and FANCI.

33 ial for the monoubiquitination of FANCD2 and FANCI.

34 not for the monoubiquitination of FANCD2 and FANCI.

35 monoubiquitinates its substrates FANCD2 and FANCI.

36 nt attachment of monoubiquitin to FANCD2 and FANCI.

37 accumulation of monoubiquitinated FANCD2 and FANCI.

38 nd chromatin localization of both FANCD2 and FANCI.

39 ion and chromatin localization of FANCD2 and FANCI.

40 ns, FA complementation group D2 (FANCD2) and FANCI.

41 , retention of monoubiquitinated FANCD2, and FANCI in chromatin, and for efficient ICL repair.

42 assembly of the FA core complex, FANCD2, and FANCI into DNA repair foci.

43 FANCL with its partners, Ube2t, FANCD2, and FANCI.

44 s, we generated isogenic FANCI-, FANCD2- and FANCI:FANCD2 double-null cells.

45 stic insight into the functions of FANCL and FANCI in the catalysis of FANCD2 monoubiquitination.

46 x monoubiquitinates and recruits the central FANCI and FANCD2 proteins that subsequently coordinate I

47 In contrast, FANCI is dispensable for FANCD2-dependent BLMcx regulati

48 cient for FA complementation group I and D2 (FANCI and FANCD2) that function as part of the FA I-D2 c

49 y is the Fanconi anemia I-Fanconi anemia D2 (FANCI-FANCD2) (ID) complex, which is activated by DNA da

50 Upon the occurrence of DNA damage, FANCI becomes monoubiquitinated on Lys-523 and relocaliz

51 Here, we focus on an FA-I patient-derived FANCI mutant protein, R1299X (deletion of 30 residues fr

52 Although loss of either FANCI or FANCD2 is known to prevent monoubiquitination o

53 hed DNA structures when compared with either FANCI or FANCD2 alone.

54 on of FA effector proteins FANCI and FANCD2 (FANCI-D2) and required the viral DNA polymerase.

55 Like FANCD2, FANCI is monoubiquitinated and unexpectedly, ubiquitinat

56 rylation is the molecular trigger for FANCD2-FANCI dissociation.

57 rst structural insight into the human FANCD2-FANCI complex by obtaining the cryo-EM structure.

58 ults uncover the mechanism of how the FANCD2-FANCI complex activates the FA pathway, and explains the

59 We show that the FANCD2-FANCI complex forms independently of ATR and FA core com

60 The FANCD2-FANCI complex is central to the pathway, and localizes t

61 egulated or what the functions of the FANCD2-FANCI complex versus the monomeric proteins are.

62 diating the monoubiquitination of the FANCD2-FANCI complex.

63 essive mutations in the gene encoding FANCD2/FANCI-associated nuclease 1 (FAN1) cause KIN in humans.

64 lear core complex to monoubiquitinate FANCD2/FANCI in response to DNA damage.

65 of their ability to monoubiquitinate FANCD2/FANCI.

66 Deficiency of FANCD2/FANCI-associated nuclease 1 (FAN1) in humans leads to ka

67 How USP1/UAF1 is targeted to the FANCD2/FANCI heterodimer has remained unknown.

68 each carried nonsense variant in the FANCD2/FANCI-associated nuclease 1 gene (FAN1), which encodes a

69 biquitination and deubiquitination of FANCD2:FANCI heterodimer is central to DNA repair in a pathway

70 s for temporal and spatial control of FANCD2:FANCI monoubiquitination that is critical for chemothera

71 nd restrict monoubiquitination to the FANCD2:FANCI heterodimer in only a DNA-bound form.

72 FANCC, FANCD1, FANCD2, FANCE, FANCF, FANCG, FANCI, FANCJ, FANCL, FANCM, and FANCN).

73 These data reveal a unique role for FANCI as a modulator of dormant origin firing and link t

74 trating functional separation of FANCD2 from FANCI.

75 ivation triggers dissociation of FANCD2 from FANCI.

76 itylation of the FA complementation group I (FANCI)-FANCD2 (ID) complex by FA complementation group L

77 To address if FANCI is also involved in these FANCD2-dependent mechani

78 Importantly, FANCI and FANCD2 monoubiquitination is co-dependent, sug

79 Mutation in FANCI is responsible for loss of a functional FA pathway

80 Interestingly, mutations in FANCI that impair its DNA binding activity compromise DN

81 aracterize important structural region(s) in FANCI that is required to activate the FA pathway.

82 -dependent mechanisms, we generated isogenic FANCI-, FANCD2- and FANCI:FANCD2 double-null cells.

83 Cells lacking FANCI have reduced number of origins, increased inter-or

84 Intriguingly, ATR-mediated FANCI phosphorylation inhibits dormant origin firing whi

85 recruited to lesions by a monoubiquitinated FANCI-FANCD2 (ID) complex and participates in ICL repair

86 Simultaneously, FANCD2-but not FANCI-plays a major role in HDR-mediated replication res

87 E motif, that is critical for the ability of FANCI to properly monoubiquitinate FANCD2 and promote DN

88 Recombination acts downstream of FANCI-FANCD2, yet RAD51 binds ICL-stalled replication fo

89 pressed in insect cells, a small fraction of FANCI forms a stable complex with FANCD2 (Fanconi anemia

90 L-stalled replication forks independently of FANCI-FANCD2 and before DSB formation.

91 hat, although proper nuclear localization of FANCI is crucial for robust FANCD2 monoubiquitination, t

92 The 7.8 angstrom electron-density map of FANCI-DNA crystals and in vitro data show that each prot

93 Like FANCD2, monoubiquitination of FANCI requires the FA core complex.

94 significantly precedes monoubiquitination of FANCI; moreover, monoubiquitination responses of FANCD2

95 An increased number of FANCI foci form and become resistant to Triton X extract

96 hway activation relies on phosphorylation of FANCI by the ataxia telangiectasia and Rad3-related (ATR

97 Although the C terminus of FANCI is dispensable for direct DNA binding, it seems to

98 on of chromatin-bound FANCD2 exceeds that of FANCI throughout replication.

99 ting chromatin prior to-and independently of-FANCI.

100 nuclear speckles in a manner that depends on FANCI and on the activity of the checkpoint kinase ATR.

101 2 sequence of UAF1 or mutation of the SIM on FANCI disrupts UAF1/FANCI binding and inhibits FANCD2 de

102 SUMO-like domain-interacting motif (SIM) on FANCI.

103 r pathway that does not require incisions or FANCI-FANCD2.

104 . now demonstrate that FANCD2 has a paralog, FANCI.

105 the Fanconi anemia (FA) DNA repair pathway, FANCI, as a key effector of dormant origin firing in res

106 A phosphodead FANCI mutant fails to dissociate from FANCD2, whereas ph

107 ssociate from FANCD2, whereas phosphomimetic FANCI cannot interact with FANCD2, indicating that FANCI

108 Here we describe the FA protein FANCI, identified as an ATM/ATR kinase substrate require

109 of either FANCD2 or another key FA protein, FANCI.

110 n monoubiquitination of FA effector proteins FANCI and FANCD2 (FANCI-D2) and required the viral DNA p

111 epair pathway, the tumor suppressor proteins FANCI and FANCD2 (the ID complex), are SUMOylated in res

112 ubiquitination of 2 interacting FA proteins, FANCI and FANCD2.

113 Intriguingly, the purified FANCI-FANCD2 complex preferentially binds to the branche

114 bust FANCD2 monoubiquitination, the putative FANCI EDGE motif is important for DNA crosslink repair.

115 TR (ataxia telangiectasia and Rad3-related), FANCI, and FANCD2.

116 n the absence of FANCD2, DNA also stimulates FANCI monoubiquitination, but in a FANCL-independent man

117 We also demonstrate that FANCI can be ubiquitinated on Lys-523 by the UBE2T-FANCL

118 We also demonstrate that FANCI forms discrete nuclear foci in HeLa cells in the a

119 tation with purified proteins indicates that FANCI interacts with FANCD2 through its C-terminal amino

120 cannot interact with FANCD2, indicating that FANCI phosphorylation is the molecular trigger for FANCD

121 We propose that FANCI and FANCD2 have partially non-overlapping and poss

122 We show that FANCI and FANCD2 are partially independent regarding the

123 Here we show that FANCI and FANCD2 associate with splicing factor 3B1 (SF3

124 We show that FANCI and its C-terminal fragment possess a DNA binding

125 ng super-resolution microscopy, we show that FANCI co-localizes with MCM-bound chromatin in response

126 Using a cell-free system, we showed that FANCI-FANCD2 is required for replication-coupled ICL rep

127 The FANCI foci are colocalized perfectly with FANCD2 and par

128 The FANCI protein associates with FANCD2 and, together, as t

129 associates with FANCD2 and, together, as the FANCI-FANCD2 (ID) complex, localize to chromatin in resp

130 At the heart of this pathway is the FANCI-FAND2 (ID) complex, which, upon ubiquitination by

131 Furthermore, addition of the FANCI protein enhances monoubiquitination and also restr

132 his pathway is the monoubiquitination of the FANCI-FANCD2 (ID) complex by the multiprotein "core comp

133 ty responsible for monoubiquitination of the FANCI-FANCD2 (ID) complex, which in turn initiates a cas

134 ia pathway is the mono-ubiquitylation of the FANCI-FANCD2 complex, but how this complex confers ICL r

135 Our data suggest that the FANCI-FANCD2 complex may participate in repair of damage

136 ain and monoubiquitin covalently attached to FANCI, and that this interaction shields monoubiquitinat

137 on forks collide with the lesion, leading to FANCI-FANCD2-dependent unhooking and formation of a doub

138 study, we report that the purified wild-type FANCI (Fanconi anemia complementation group I) protein d

139 r mutation of the SIM on FANCI disrupts UAF1/FANCI binding and inhibits FANCD2 deubiquitination and D

140 rsal of these defects requires ubiquitylated FANCI-FANCD2.

141 cleavage is prevented, unhooking occurs via FANCI-FANCD2-dependent incisions.

142 ts respective partner, it is unclear whether FANCI has any additional domains that may be important i

143 CUE domain is required for interaction with FANCI, retention of monoubiquitinated FANCD2, and FANCI