Protection of hybridoma cells against apoptosis by a loop domain-deficient Bcl-xL protein |
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Authors: | Joel Charbonneau Eric Gauthier |
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Institution: | (1) Cellular Biochemistry Research Laboratory , Department of Chemistry and Biochemistry, Laurentian University, Canada;(2) Department of Chemistry and Biochemistry, Laurentian University, 935 Ramsey Lake Road, Sudbury, Ontario, P 3E 2C 6, Canada |
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Abstract: | The ectopic expression of several members of the Bcl-2 family of anti-apoptotic proteins is a promising strategy to improve
the viability of hybridoma cells in culture. However, the impact of post-translational modifications on the function of these
proteins in murine hybridomas is unknown. To address this issue, the anti-apoptotic properties of a mutant of Bcl-xL devoid
of the so-called “loop domain„ (Bcl-xL▵ 46-83) were investigated using the Sp2/ O-Ag14 hybridoma model. Clones of Sp2/ O-Ag14
cells expressing Bcl-xL▵ 46-83 exhibited resistance against L-glutamine deprivation to similar levels than cells expressing
the wild type protein. In contrast, protection against the cytotoxic effects of cycloheximide (CHX) was highly dependent on
the level of expression of the Bcl-xL▵ 46-83 mutant. Analysis of the growth behaviour of the transfected cells showed that
Bcl-xL▵ 46-83 was superior to the wild type protein in prolonging Sp2/ O-Agl4 cell viability in stationary batch culture.
Furthermore, the prolongation of cell viability in batch culture was directly proportional to the level of expression of the
mutated protein. Our results indicate that removal of the loop domain improves the anti-apoptotic activity of Bcl-xL in hybridoma
cells grown in stationary batch culture.
This revised version was published online in August 2006 with corrections to the Cover Date. |
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Keywords: | Apoptosis Bcl-xL Cell viability Hybridoma Loop domain Mutagenesis |
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