Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2010 Mar;17(3):294-8.
doi: 10.1038/nsmb.1759. Epub 2010 Feb 14.

The structure of the catalytic subunit FANCL of the Fanconi anemia core complex

Ambrose R Cole  1 , Laurence P C Lewis, Helen Walden
Affiliations

The structure of the catalytic subunit FANCL of the Fanconi anemia core complex

Ambrose R Cole et al. Nat Struct Mol Biol. 2010 Mar.

Abstract

The Fanconi anemia (FA) pathway is activated in response to DNA damage, leading to monoubiquitination of the substrates FANCI and FANCD2 by the FA core complex. Here we report the crystal structure of FANCL, the catalytic subunit of the FA core complex, at 3.2 A. The structure reveals an architecture fundamentally different from previous sequence-based predictions. The molecule is composed of an N-terminal E2-like fold, which we term the ELF domain, a novel double-RWD (DRWD) domain, and a C-terminal really interesting new gene (RING) domain predicted to facilitate E2 binding. Binding assays show that the DRWD domain, but not the ELF domain, is responsible for substrate binding.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Domain architecture of FANCL
a. Ribbon diagram depicting the FANCL structure. The ELF, DRWD and RING domains are coloured red, yellow/lime and blue, respectively. Zinc atoms are represented by grey spheres and secondary structure elements are labelled. Two views are displayed related by a 90° rotation around the x-axis, as indicated. b. Surface representation of FANCL coloured by sequence conservation between divergent eukaryotic species. Grey indicates non-conserved residues, salmon semi-conservative substitutions, orange conservative substitutions and red conserved residues, shown in the alignment in Supplementary figure 2.
Figure 2
Figure 2. Comparison of UBC-superfamily folds with E2s and RWD proteins
A catalytically active E2 (UbcH7) is shown in cyan, the inactive E2 MMS2 in blue, and RWD2-2 in magenta on the top row (pdb codes 1fbv, 1j7d and 2daw, respectively). The catalytic cysteine of UbcH7 is represented as a sphere, structural features are indicated, and known protein–protein interaction surfaces are highlighted in pink. The bottom row shows the ELF domain in red, and the N-terminal and C-terminal lobes of the DRWD domain in yellow and lime, respectively. These folds are in the same orientation as the UBC-superfamily members, and protein–protein interaction surfaces are shaded in magenta. Residues highlighted in the structural analysis are coloured cyan.
Figure 3
Figure 3. Protein–protein interaction surfaces on FANCL
a. The ELF domain has 3 exposed hydrophobic patches coloured purple, pink and blue. Two views related by a 90° rotation around the x-axis are shown, represented as both surface patches and ball-and-stick residues. Individual residues are labelled. b. The DRWD domain has 3 hydrophobic patches shown in salmon, green and cyan. The view is 180° around the x-axis in relation to the left-hand panel in Figure 1a, represented as both surface patches and ball-and-stick residues. Individual residues are labelled. c. The RING domain coordinates 2 zinc atoms represented by grey spheres. Coordinating residues are labelled. d. The DRWD–RING interface. DRWD residues are indicated in lime, RING residues are in blue, and individual residues are labelled.
Figure 4
Figure 4. The DRWD domain recruits substrate
a. Pulldown binding assays using His-SUMOSTAR-FANCD2(1–762) as bait. Input samples (I) of FANCL species and samples bound by His-SUMOSTAR-FANCD2(1–762) (B) are indicated. Samples are analysed by Western blotting using anti-FANCL or anti-SUMO antibodies. b. Pulldown binding assays using His-SUMOSTAR-FANCI as bait. Input samples (I) of FANCL species and samples bound by His-SUMOSTAR-FANCI (B) are indicated. Samples are analysed by Western blotting using anti-FANCL and anti-SUMO antibodies. c. A schematic representation of FANCD2 monoubiquitination. The I/D2 complex is represented in green, with Lys561 indicated. The ELF, DRWD and RING domains are red, yellow and blue, respectively. The E2 and ubiquitin (U) are indicated.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Alter BP. Fanconi's anemia and malignancies. Am J Hematol. 1996;53:99–110. - PubMed
    1. Howlett NG, et al. Biallelic inactivation of BRCA2 in Fanconi anemia. Science. 2002;297:606–609. - PubMed
    1. Strathdee CA, Duncan AM, Buchwald M. Evidence for at least four Fanconi anaemia genes including FACC on chromosome 9. Nat Genet. 1992;1:196–198. - PubMed
    1. Lo Ten Foe JR, et al. Expression cloning of a cDNA for the major Fanconi anaemia gene, FAA. Nat Genet. 1996;14:320–323. - PubMed
    1. Meetei AR, et al. X-linked inheritance of Fanconi anemia complementation group B. Nat Genet. 2004;36:1219–1224. - PubMed

Publication types

MeSH terms

Substances

Associated data