Knockdown of alphaII spectrin in normal human cells by siRNA leads to chromosomal instability and decreased DNA interstrand cross-link repair.
Nonerythroid alpha-spectrin (alphaIISp) is a structural protein involved in repair of DNA interstrand cross-links and is deficient in cells from patients with Fanconi anemia (FA), which are defective in ability to repair cross-links. In order to further demonstrate the importance of the role that alphaIISp plays in normal human cells and in the repair defect in FA, alphaIISp was knocked down in normal cells using siRNA. Depletion of alphaIISp in normal cells by siRNA resulted in chromosomal instability and cellular hypersensitivity to DNA interstrand cross-linking agents. An increased number of chromosomal aberrations were observed and, following treatment with a DNA interstrand cross-linking agent, mitomycin C, cells showed decreased cell growth and survival and decreased formation of damage-induced alphaIISp and XPF nuclear foci. Thus depletion of alphaIISp in normal cells leads to a number of defects observed in FA cells, such as chromosome instability and a deficiency in cross-link repair.
McMahon
Laura W
LW
Department of Pathology and Laboratory Medicine, UMDNJ-New Jersey Medical School and the Graduate School of Biomedical Sciences, 185 South Orange Avenue, Newark, NJ 07103, USA.
Zhang
Pan
P
Sridharan
Deepa M
DM
Lefferts
Joel A
JA
Lambert
Muriel W
MW
D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D043171:Chromosomal Instability / Q000235:genetics*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000235:genetics; D004260:DNA Repair / Q000235:genetics*; D005199:Fanconi Anemia / Q000235:genetics*; D055785:Gene Knockdown Techniques; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000502:physiology*; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D013049:Spectrin
D002352:Carrier Proteins; D003432:Cross-Linking Reagents; D004247:DNA; D008840:Microfilament Proteins; D034741:RNA, Small Interfering; D013049:Spectrin