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Deconstructing Survivin: comprehensive genetic analysis of Survivin function by conditional knockout in a vertebrate cell line
- Source :
- The Journal of Cell Biology, Yue, Z, Carvalho, A, Xu, Z, Yuan, X, Cardinale, S, Ribeiro, S, Lai, F, Ogawa, H, Gudmundsdottir, E, Gassmann, R, Morrison, C G, Ruchaud, S & Earnshaw, W C 2008, ' Deconstructing Survivin: comprehensive genetic analysis of Survivin function by conditional knockout in a vertebrate cell line ', Journal of Cell Biology, vol. 183, no. 2, pp. 279-296 . https://doi.org/10.1083/jcb.200806118
- Publication Year :
- 2008
-
Abstract
- Survivin is a key cellular protein thought to function in apoptotic regulation, mitotic progression, or possibly both. In this study, we describe the isolation of two conditional knockouts of the survivin gene in chicken DT40 cells. DT40 cells lacking Survivin die in interphase after failing to complete cytokinesis. However, these cells show normal sensitivity to the chemotherapeutic agent etoposide. Expression of Survivin mutants against a null background to reassess the role of several key residues reveals that DT40 cells can grow normally if their sole Survivin is missing a widely studied cyclin-dependent kinase phosphorylation site or sites reportedly essential for binding to Smac or aurora B. Mutations in the nuclear export sequence or dimerization interface render cells temperature sensitive for growth. As an important caveat for other studies in which protein function is studied by transient transfection, three of the Survivin mutants fail to localize in the presence of the wild-type protein but do localize and indeed support life in its absence.
- Subjects :
- Amino Acid Motifs
Cell Separation
medicine.disease_cause
0302 clinical medicine
Aurora Kinases
Conditional gene knockout
Phosphorylation
Research Articles
Etoposide
0303 health sciences
Mutation
biology
Cell Death
Temperature
Cyclin-Dependent Kinases
Cell biology
Phenotype
Phosphothreonine
030220 oncology & carcinogenesis
Vertebrates
Microtubule-Associated Proteins
Protein Binding
Cell Survival
Centromere
Mitosis
Spindle Apparatus
Protein Serine-Threonine Kinases
Article
Cell Line
Mitochondrial Proteins
03 medical and health sciences
Cyclin-dependent kinase
Survivin
medicine
Animals
Nuclear export signal
Gene knockout
030304 developmental biology
Cytokinesis
Aspartic Acid
Cell Biology
Staurosporine
Molecular biology
Protein Structure, Tertiary
biology.protein
Chickens
Subjects
Details
- ISSN :
- 15408140
- Volume :
- 183
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- The Journal of cell biology
- Accession number :
- edsair.doi.dedup.....bec0801320c910c95f2e1796e7b5d643