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Mutations of the SLX4 gene in Fanconi anemia.

Kim Y, Lach FP, Desetty R, Hanenberg H, Auerbach AD, Smogorzewska A - Nat. Genet. (2011)

Bottom Line: Fanconi anemia is a rare recessive disorder characterized by genome instability, congenital malformations, progressive bone marrow failure and predisposition to hematologic malignancies and solid tumors.Depletion of SLX4, which interacts with multiple nucleases and has been recently identified as a Holliday junction resolvase, results in increased sensitivity of the cells to DNA crosslinking agents.Here we report the identification of biallelic SLX4 mutations in two individuals with typical clinical features of Fanconi anemia and show that the cellular defects in these individuals' cells are complemented by wildtype SLX4, demonstrating that biallelic mutations in SLX4 (renamed here as FANCP) cause a new subtype of Fanconi anemia, Fanconi anemia-P.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, New York, USA.

ABSTRACT
Fanconi anemia is a rare recessive disorder characterized by genome instability, congenital malformations, progressive bone marrow failure and predisposition to hematologic malignancies and solid tumors. At the cellular level, hypersensitivity to DNA interstrand crosslinks is the defining feature in Fanconi anemia. Mutations in thirteen distinct Fanconi anemia genes have been shown to interfere with the DNA-replication-dependent repair of lesions involving crosslinked DNA at stalled replication forks. Depletion of SLX4, which interacts with multiple nucleases and has been recently identified as a Holliday junction resolvase, results in increased sensitivity of the cells to DNA crosslinking agents. Here we report the identification of biallelic SLX4 mutations in two individuals with typical clinical features of Fanconi anemia and show that the cellular defects in these individuals' cells are complemented by wildtype SLX4, demonstrating that biallelic mutations in SLX4 (renamed here as FANCP) cause a new subtype of Fanconi anemia, Fanconi anemia-P.

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SLX4 is defective in two FA patients. A. Schematic of SLX4 (based on ref. 7) showing the domain architecture, the interacting proteins, and the predicted protein effect of SLX4 mutations in IFAR1084/1 and IFAR414/1 patients. B. Analysis of the mutant SLX4 protein in the patient’s cell lines. Cell extracts of primary BJ, RA3083, and RA3331 fibroblasts were subjected to immunoprecipitation using a control rabbit antibody (control IgG) or the SLX4 antibody. Asterisks indicate the crossreacting bands. Note that the antibody does not identify SLX4 in straight western (lanes 7 to 9).
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Figure 2: SLX4 is defective in two FA patients. A. Schematic of SLX4 (based on ref. 7) showing the domain architecture, the interacting proteins, and the predicted protein effect of SLX4 mutations in IFAR1084/1 and IFAR414/1 patients. B. Analysis of the mutant SLX4 protein in the patient’s cell lines. Cell extracts of primary BJ, RA3083, and RA3331 fibroblasts were subjected to immunoprecipitation using a control rabbit antibody (control IgG) or the SLX4 antibody. Asterisks indicate the crossreacting bands. Note that the antibody does not identify SLX4 in straight western (lanes 7 to 9).

Mentions: Sequencing of the cDNA from the 1084/1 patient’s cells revealed skipping of Exon 5 (Figure S2A), due to a homozygous point mutation in the canonical splice donor dinucleotide GT in intron 5 (c.1163+2T>A) in the genomic DNA (Figure S2B). Both parents were found to be heterozygous and an unaffected sibling was found to be negative for this mutation (Figure S2B). The predicted effect of this mutation is a 70 amino acid deletion of amino acids (aa) 317 to 387 of SLX4 (p.R317_F387del), leading to an in-frame deletion of the conserved Cys and Leu of the first UBZ domain and the whole second UBZ domain (Figure 2A, Figure S2C). Immunoprecipitation of SLX4 from the cell line RA3083 confirmed the presence of a slightly shorter protein product (Figure 2B, lane 5, Figure S2D)


Mutations of the SLX4 gene in Fanconi anemia.

Kim Y, Lach FP, Desetty R, Hanenberg H, Auerbach AD, Smogorzewska A - Nat. Genet. (2011)

SLX4 is defective in two FA patients. A. Schematic of SLX4 (based on ref. 7) showing the domain architecture, the interacting proteins, and the predicted protein effect of SLX4 mutations in IFAR1084/1 and IFAR414/1 patients. B. Analysis of the mutant SLX4 protein in the patient’s cell lines. Cell extracts of primary BJ, RA3083, and RA3331 fibroblasts were subjected to immunoprecipitation using a control rabbit antibody (control IgG) or the SLX4 antibody. Asterisks indicate the crossreacting bands. Note that the antibody does not identify SLX4 in straight western (lanes 7 to 9).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3345287&req=5

Figure 2: SLX4 is defective in two FA patients. A. Schematic of SLX4 (based on ref. 7) showing the domain architecture, the interacting proteins, and the predicted protein effect of SLX4 mutations in IFAR1084/1 and IFAR414/1 patients. B. Analysis of the mutant SLX4 protein in the patient’s cell lines. Cell extracts of primary BJ, RA3083, and RA3331 fibroblasts were subjected to immunoprecipitation using a control rabbit antibody (control IgG) or the SLX4 antibody. Asterisks indicate the crossreacting bands. Note that the antibody does not identify SLX4 in straight western (lanes 7 to 9).
Mentions: Sequencing of the cDNA from the 1084/1 patient’s cells revealed skipping of Exon 5 (Figure S2A), due to a homozygous point mutation in the canonical splice donor dinucleotide GT in intron 5 (c.1163+2T>A) in the genomic DNA (Figure S2B). Both parents were found to be heterozygous and an unaffected sibling was found to be negative for this mutation (Figure S2B). The predicted effect of this mutation is a 70 amino acid deletion of amino acids (aa) 317 to 387 of SLX4 (p.R317_F387del), leading to an in-frame deletion of the conserved Cys and Leu of the first UBZ domain and the whole second UBZ domain (Figure 2A, Figure S2C). Immunoprecipitation of SLX4 from the cell line RA3083 confirmed the presence of a slightly shorter protein product (Figure 2B, lane 5, Figure S2D)

Bottom Line: Fanconi anemia is a rare recessive disorder characterized by genome instability, congenital malformations, progressive bone marrow failure and predisposition to hematologic malignancies and solid tumors.Depletion of SLX4, which interacts with multiple nucleases and has been recently identified as a Holliday junction resolvase, results in increased sensitivity of the cells to DNA crosslinking agents.Here we report the identification of biallelic SLX4 mutations in two individuals with typical clinical features of Fanconi anemia and show that the cellular defects in these individuals' cells are complemented by wildtype SLX4, demonstrating that biallelic mutations in SLX4 (renamed here as FANCP) cause a new subtype of Fanconi anemia, Fanconi anemia-P.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Genome Maintenance, The Rockefeller University, New York, New York, USA.

ABSTRACT
Fanconi anemia is a rare recessive disorder characterized by genome instability, congenital malformations, progressive bone marrow failure and predisposition to hematologic malignancies and solid tumors. At the cellular level, hypersensitivity to DNA interstrand crosslinks is the defining feature in Fanconi anemia. Mutations in thirteen distinct Fanconi anemia genes have been shown to interfere with the DNA-replication-dependent repair of lesions involving crosslinked DNA at stalled replication forks. Depletion of SLX4, which interacts with multiple nucleases and has been recently identified as a Holliday junction resolvase, results in increased sensitivity of the cells to DNA crosslinking agents. Here we report the identification of biallelic SLX4 mutations in two individuals with typical clinical features of Fanconi anemia and show that the cellular defects in these individuals' cells are complemented by wildtype SLX4, demonstrating that biallelic mutations in SLX4 (renamed here as FANCP) cause a new subtype of Fanconi anemia, Fanconi anemia-P.

Show MeSH
Related in: MedlinePlus