BRCA1-independent ubiquitination of FANCD2

Cassandra J. Vandenberg; Fanni Gergely; Chong Yi Ong; Paul Pace; Donna L. Mallery; Kevin Hiom; Ketan J. Patel

doi:10.1016/S1097-2765(03)00281-8

BRCA1-independent ubiquitination of FANCD2

Cassandra J. Vandenberg, Fanni Gergely, Chong Yi Ong, Paul Pace, Donna L. Mallery, Kevin Hiom (Lead / Corresponding author), Ketan J. Patel (Lead / Corresponding author)

Research output: Contribution to journal › Article › peer-review

122 Citations (Scopus)

Abstract

Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

Original language	English
Pages (from-to)	247-254
Number of pages	8
Journal	Molecular Cell
Volume	12
Issue number	1
DOIs	https://doi.org/10.1016/S1097-2765(03)00281-8
Publication status	Published - Jul 2003

Keywords

Animals
BRCA1 protein
Carrier proteins
Cell-free system
DNA damage
Fanconi anemia
Fanconi anemia complementation group D2 protein
Gene expression regulation, Neoplastic
HeLa cells
Humans
Iron-binding proteins
Ligases
Mutation
Nuclear proteins
Protein structure, Tertiary
RNA, Small interfering
Tumor suppressor proteins
Ubiquitin
Ubiquitin-conjugating Enzymes
Ubiquitin-protein ligases

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title = "BRCA1-independent ubiquitination of FANCD2",

abstract = "Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.",

keywords = "Animals, BRCA1 protein, Carrier proteins, Cell-free system, DNA damage, Fanconi anemia, Fanconi anemia complementation group D2 protein, Gene expression regulation, Neoplastic, HeLa cells, Humans, Iron-binding proteins, Ligases, Mutation, Nuclear proteins, Protein structure, Tertiary, RNA, Small interfering, Tumor suppressor proteins, Ubiquitin, Ubiquitin-conjugating Enzymes, Ubiquitin-protein ligases",

author = "Vandenberg, {Cassandra J.} and Fanni Gergely and Ong, {Chong Yi} and Paul Pace and Mallery, {Donna L.} and Kevin Hiom and Patel, {Ketan J.}",

year = "2003",

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doi = "10.1016/S1097-2765(03)00281-8",

language = "English",

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T1 - BRCA1-independent ubiquitination of FANCD2

AU - Vandenberg, Cassandra J.

AU - Gergely, Fanni

AU - Ong, Chong Yi

AU - Pace, Paul

AU - Mallery, Donna L.

AU - Hiom, Kevin

AU - Patel, Ketan J.

PY - 2003/7

Y1 - 2003/7

N2 - Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

AB - Monoubiquitination of the FANCD2 protein is a key step in the Fanconi anemia (FA) tumor suppressor pathway, coinciding with this molecule's accumulation at sites of genome damage. Strong circumstantial evidence points to a requirement for the BRCA1 gene product in this step. Here, we show that the purified BRCA1/BARD1 complex, together with E1 and UbcH5a, is sufficient to reconstitute the monoubiquitination of FANCD2 in vitro. Although siRNA-mediated knockdown of BRCA1 in human cells results in defective targeting of FANCD2 to sites of DNA damage, it does not lead to a defect in FANCD2 ubiquitination. Furthermore, ablation of the RING finger domains of either BRCA1 or BARD1 in the chicken B cell line DT40 also leaves FANCD2 modification intact. Consequently, while BRCA1 affects the accumulation of FANCD2 at sites of DNA damage, BRCA1/BARD1 E3 ligase activity is not essential for the monoubiquitination of FANCD2.

KW - Animals

KW - BRCA1 protein

KW - Carrier proteins

KW - Cell-free system

KW - DNA damage

KW - Fanconi anemia

KW - Fanconi anemia complementation group D2 protein

KW - Gene expression regulation, Neoplastic

KW - HeLa cells

KW - Humans

KW - Iron-binding proteins

KW - Ligases

KW - Mutation

KW - Nuclear proteins

KW - Protein structure, Tertiary

KW - RNA, Small interfering

KW - Tumor suppressor proteins

KW - Ubiquitin

KW - Ubiquitin-conjugating Enzymes

KW - Ubiquitin-protein ligases

U2 - 10.1016/S1097-2765(03)00281-8

DO - 10.1016/S1097-2765(03)00281-8

M3 - Article

C2 - 12887909

SN - 1097-2765

VL - 12

SP - 247

EP - 254

JO - Molecular Cell

JF - Molecular Cell

IS - 1

ER -

BRCA1-independent ubiquitination of FANCD2

Abstract

Keywords

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