Effect of mutations of murine lens αB crystallin on transfected neural cell viability and cellular translocation in response to stress

FEBS Letters

Coop, Audrey¹  Crabbe, M.James C¹  Goode, Derek¹  Hepburne-Scott, Henry W¹  Wiesmann, Kirsten E.H¹ 

[1] Division of Cell and Molecular Biology, School of Animal and Microbial Sciences, The University of Reading, P.O. Box 228, Whiteknights, Reading, Berks. RG6 6AJ, UK

关键词: Chaperone;    Neurological disorder;    Cataract;    Neuroblastoma;    Homeobox;   

DOI  :  10.1016/S0014-5793(98)01260-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We examined the influence of over-expressed native and mutant murine lens αB crystallin on the response of a murine neural cell line to heat and ionic strength shock. Native and mutant (F27R and KK174/175LL) murine αB crystallin amplicons were subcloned into a Lac-Switch IPTG-inducible RSV promoter eukaryotic vector, and transfected into N1E-115 cells using lipofectin. Expression was induced maximally 8 h after addition of IPTG (optimal final concentration 1 mM) to the medium. Cells grew normally after transfection with native and mutant murine αB crystallin. We demonstrated expression of the protein using specific anti-α crystallin antibodies. Viability under severe heat and ionic strength stress increased when cells had been transfected with native αB crystallin, but not with mutants F27R or KK174/175LL. Heat shock caused translocation of both native and mutant αB crystallins into the central region of the cells. These results show that mutations in αB crystallin that effect its chaperone-like activity may also influence viability of N1E-115 neural cells under stress, while not influencing the distribution of the protein within the cell.

【 授权许可】

Unknown