Human alpha spectrin h and the fanconi anemia proteins fanca, fancc and fancg, bind to dna containing interstrand cross-links

M. W. Lambert, J. Sangerman, Steven Goodman, L. W. McMahon

Research output: Contribution to journalArticle

Abstract

Fanconi anemia (FA) is a genetic disorder characterized by bone marrow failurb, congenital anomalies, cancer susceptibility and a cellular hypersensitivity to DNA interstrand cross-linking agents which correlates with a defect in ability to repair this type of damage. We have recently identified a spectrin II (αSplI∑*) as a component of a nuclear protein complex that is involved in repair of DNA interstrand cross-links, aid shown that it forms a complex with the FANCA, FANCC and FANCG proteins and is deficient in FA-A, FA-B, FA-C, FA-D, and FA-G cells. The ability of αSpll∑* and the FA proteins to bind to DNA containing interstrand cross-links has now been examined. Using DNA affinity chromatography, chromatin-associated protein extracts from HeLa cells were examined for proteins which bound to a 105 bp oligonucleotide that contained 4, i1, 8-trimethylpsoralen interstrand cross-links. Immunoblot analysis showed that aSpltëf bound to the cross-linked DNA as did the FANCA, FANCC, and FANCG proteinls. Competition experiments indicated that aSplE had binding specificity for cross-linked DNA. Further analysis showed that purified bovine brain spectrin (aSpllZl/βSpllZl)2 bound directly to the damaged DNA. Whether the FA proteins bind directly to the crosslinked DNA or indirectly via their ability to form a complex with aSpllZ is und :r investigation. aSpllS could thus play a role in cross-link repair via its affinity for crosslinked DNA. It could bind to the damaged DNA and act as a scaffold to help target repdr proteins to the site of damage and help align or enhance interactions between them ar d between the FA proteins. A deficiency in aSpllZ in FA cells could thus reduce the efficiency of the repair process in these cells, as has been observed. It could be the damag :recognition protein we have previously shown to be defective in FA-A cells. The FA proteins could also be involved in the damage-recognition step or, possibly, due to their interaction with ctSpllE, in some other aspect of the DNA repair process or in some other cellular events. Since spectrin plays a role in a number of different cellular processes, such as signal transduction and cell growth and differentiation, a deficiency in this prote n could affect a number of systems in FA cells, possibly accounting for some of the diver ;e cellular and clinical defects that have been observed in FA.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART I
StatePublished - 2000
Externally publishedYes

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Fanconi Anemia Complementation Group Proteins
Fanconi Anemia
Spectrin
DNA
Repair
Fanconi Anemia Complementation Group G Protein
Fanconi Anemia Complementation Group A Protein
DNA Repair
Fanconi Anemia Complementation Group C Protein
Proteins
Trioxsalen
Gastrin-Secreting Cells
Inborn Genetic Diseases
Affinity chromatography
Signal transduction
Defects
Nuclear Proteins
Affinity Chromatography
HeLa Cells
Cell growth

All Science Journal Classification (ASJC) codes

  • Hematology

Cite this

Human alpha spectrin h and the fanconi anemia proteins fanca, fancc and fancg, bind to dna containing interstrand cross-links. / Lambert, M. W.; Sangerman, J.; Goodman, Steven; McMahon, L. W.

In: Blood, Vol. 96, No. 11 PART I, 2000.

Research output: Contribution to journalArticle

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abstract = "Fanconi anemia (FA) is a genetic disorder characterized by bone marrow failurb, congenital anomalies, cancer susceptibility and a cellular hypersensitivity to DNA interstrand cross-linking agents which correlates with a defect in ability to repair this type of damage. We have recently identified a spectrin II (αSplI∑*) as a component of a nuclear protein complex that is involved in repair of DNA interstrand cross-links, aid shown that it forms a complex with the FANCA, FANCC and FANCG proteins and is deficient in FA-A, FA-B, FA-C, FA-D, and FA-G cells. The ability of αSpll∑* and the FA proteins to bind to DNA containing interstrand cross-links has now been examined. Using DNA affinity chromatography, chromatin-associated protein extracts from HeLa cells were examined for proteins which bound to a 105 bp oligonucleotide that contained 4, i1, 8-trimethylpsoralen interstrand cross-links. Immunoblot analysis showed that aSplt{\"e}f bound to the cross-linked DNA as did the FANCA, FANCC, and FANCG proteinls. Competition experiments indicated that aSplE had binding specificity for cross-linked DNA. Further analysis showed that purified bovine brain spectrin (aSpllZl/βSpllZl)2 bound directly to the damaged DNA. Whether the FA proteins bind directly to the crosslinked DNA or indirectly via their ability to form a complex with aSpllZ is und :r investigation. aSpllS could thus play a role in cross-link repair via its affinity for crosslinked DNA. It could bind to the damaged DNA and act as a scaffold to help target repdr proteins to the site of damage and help align or enhance interactions between them ar d between the FA proteins. A deficiency in aSpllZ in FA cells could thus reduce the efficiency of the repair process in these cells, as has been observed. It could be the damag :recognition protein we have previously shown to be defective in FA-A cells. The FA proteins could also be involved in the damage-recognition step or, possibly, due to their interaction with ctSpllE, in some other aspect of the DNA repair process or in some other cellular events. Since spectrin plays a role in a number of different cellular processes, such as signal transduction and cell growth and differentiation, a deficiency in this prote n could affect a number of systems in FA cells, possibly accounting for some of the diver ;e cellular and clinical defects that have been observed in FA.",
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T1 - Human alpha spectrin h and the fanconi anemia proteins fanca, fancc and fancg, bind to dna containing interstrand cross-links

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AU - Sangerman, J.

AU - Goodman, Steven

AU - McMahon, L. W.

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N2 - Fanconi anemia (FA) is a genetic disorder characterized by bone marrow failurb, congenital anomalies, cancer susceptibility and a cellular hypersensitivity to DNA interstrand cross-linking agents which correlates with a defect in ability to repair this type of damage. We have recently identified a spectrin II (αSplI∑*) as a component of a nuclear protein complex that is involved in repair of DNA interstrand cross-links, aid shown that it forms a complex with the FANCA, FANCC and FANCG proteins and is deficient in FA-A, FA-B, FA-C, FA-D, and FA-G cells. The ability of αSpll∑* and the FA proteins to bind to DNA containing interstrand cross-links has now been examined. Using DNA affinity chromatography, chromatin-associated protein extracts from HeLa cells were examined for proteins which bound to a 105 bp oligonucleotide that contained 4, i1, 8-trimethylpsoralen interstrand cross-links. Immunoblot analysis showed that aSpltëf bound to the cross-linked DNA as did the FANCA, FANCC, and FANCG proteinls. Competition experiments indicated that aSplE had binding specificity for cross-linked DNA. Further analysis showed that purified bovine brain spectrin (aSpllZl/βSpllZl)2 bound directly to the damaged DNA. Whether the FA proteins bind directly to the crosslinked DNA or indirectly via their ability to form a complex with aSpllZ is und :r investigation. aSpllS could thus play a role in cross-link repair via its affinity for crosslinked DNA. It could bind to the damaged DNA and act as a scaffold to help target repdr proteins to the site of damage and help align or enhance interactions between them ar d between the FA proteins. A deficiency in aSpllZ in FA cells could thus reduce the efficiency of the repair process in these cells, as has been observed. It could be the damag :recognition protein we have previously shown to be defective in FA-A cells. The FA proteins could also be involved in the damage-recognition step or, possibly, due to their interaction with ctSpllE, in some other aspect of the DNA repair process or in some other cellular events. Since spectrin plays a role in a number of different cellular processes, such as signal transduction and cell growth and differentiation, a deficiency in this prote n could affect a number of systems in FA cells, possibly accounting for some of the diver ;e cellular and clinical defects that have been observed in FA.

AB - Fanconi anemia (FA) is a genetic disorder characterized by bone marrow failurb, congenital anomalies, cancer susceptibility and a cellular hypersensitivity to DNA interstrand cross-linking agents which correlates with a defect in ability to repair this type of damage. We have recently identified a spectrin II (αSplI∑*) as a component of a nuclear protein complex that is involved in repair of DNA interstrand cross-links, aid shown that it forms a complex with the FANCA, FANCC and FANCG proteins and is deficient in FA-A, FA-B, FA-C, FA-D, and FA-G cells. The ability of αSpll∑* and the FA proteins to bind to DNA containing interstrand cross-links has now been examined. Using DNA affinity chromatography, chromatin-associated protein extracts from HeLa cells were examined for proteins which bound to a 105 bp oligonucleotide that contained 4, i1, 8-trimethylpsoralen interstrand cross-links. Immunoblot analysis showed that aSpltëf bound to the cross-linked DNA as did the FANCA, FANCC, and FANCG proteinls. Competition experiments indicated that aSplE had binding specificity for cross-linked DNA. Further analysis showed that purified bovine brain spectrin (aSpllZl/βSpllZl)2 bound directly to the damaged DNA. Whether the FA proteins bind directly to the crosslinked DNA or indirectly via their ability to form a complex with aSpllZ is und :r investigation. aSpllS could thus play a role in cross-link repair via its affinity for crosslinked DNA. It could bind to the damaged DNA and act as a scaffold to help target repdr proteins to the site of damage and help align or enhance interactions between them ar d between the FA proteins. A deficiency in aSpllZ in FA cells could thus reduce the efficiency of the repair process in these cells, as has been observed. It could be the damag :recognition protein we have previously shown to be defective in FA-A cells. The FA proteins could also be involved in the damage-recognition step or, possibly, due to their interaction with ctSpllE, in some other aspect of the DNA repair process or in some other cellular events. Since spectrin plays a role in a number of different cellular processes, such as signal transduction and cell growth and differentiation, a deficiency in this prote n could affect a number of systems in FA cells, possibly accounting for some of the diver ;e cellular and clinical defects that have been observed in FA.

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