Selection of a lysine-resistant CHO-K1 mutant with reduced amino acid transport through multiple systems

High levels of L-lysine were used to select for resistant variants of Chinese hamster ovary (CHO-K1) cells. Surviving colonies were screened for altered lysine transport and two with reduced uptake were picked. Clone CH-Kr, derived from the more severely affected colony, was analyzed in detail. In s...

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Bibliographic Details
Published in:Somatic cell and molecular genetics 1993-07, Vol.19 (4), p.331-345
Main Authors: DEBUSK, W. E, ASH, J. F
Format: Article
Language:English
Subjects:
Amino Acids - metabolism
Animals
Biological and medical sciences
Biological Transport - genetics
Cell Division
Cell physiology
CHO Cells
Cloning, Molecular
Cricetinae
Drug Resistance - genetics
Fundamental and applied biological sciences. Psychology
Kinetics
Lysine - metabolism
Lysine - pharmacology
Membrane and intracellular transports
Molecular and cellular biology
Mutation
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Summary:High levels of L-lysine were used to select for resistant variants of Chinese hamster ovary (CHO-K1) cells. Surviving colonies were screened for altered lysine transport and two with reduced uptake were picked. Clone CH-Kr, derived from the more severely affected colony, was analyzed in detail. In starved cells the Vmax of lysine uptake in CH-Kr was half that of CHO while Km was unaltered. The intracellular pool of lysine, a substrate of cationic amino acid transport system y+, was significantly lower in CH-Kr. However, transport and pools of other amino acids, which are not substrates of y+, were also reduced in CH-Kr, as was the internal sodium concentration, while hexose import was increased. It appears that the mutation in CH-Kr is pleiotropic, affecting some general aspects of amino acid transport.
ISSN:0740-7750
1572-9931
DOI:10.1007/BF01232746