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Effect of mutations of murine lens αB crystallin on transfected neural cell viability and cellular translocation in response to stress
Author(s) -
Wiesmann Kirsten E.H,
Coop Audrey,
Goode Derek,
Hepburne-Scott Henry W,
Crabbe M.James C
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)01260-5
Subject(s) - transfection , mutant , microbiology and biotechnology , crystallin , heat shock protein , biology , viability assay , lac operon , chaperone (clinical) , chromosomal translocation , cell , cell culture , recombinant dna , biochemistry , gene , genetics , medicine , pathology
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.