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2. Expression of pICln in Escherichia coli gives a strong tolerance to hypotonic stress

Author

Akira Kobayashi, Guo-Zhong Tao, Yohtalou Tashima, and Hideaki Itoh

Subjects
Channel regulator, Hypotonic tolerance, Molecular Sequence Data, Biophysics, Adaptation, Biological, Expression, Biology, medicine.disease_cause, Biochemistry, Plasmid, Hypotonic Stress, Gentamicin protection assay, Structural Biology, Chloride Channels, Complementary DNA, Putative volume (or osmo)-sensitive Cl− channel, Genetics, medicine, Extracellular, Escherichia coli, Animals, Amino Acid Sequence, Cloning, Molecular, Molecular Biology, Peptide sequence, Regulation of gene expression, Base Sequence, Cell Biology, Gene Expression Regulation, Bacterial, Molecular biology, Rats, Hypotonic Solutions, pICln protein, Plasmids
Abstract

We amplified the coding region DNA sequence from a rat renal pI(Cln) cDNA by PCR and expressed the protein in Escherichia coli cells. The cells were exposed to hypotonic conditions followed by spreading them onto LB plates for subsequent colony survival assay. The present study demonstrated that the cells expressing pI(Cln) exhibit a strong resistance to hypotonic stress. Moreover, the resistance was specifically inhibited by extracellular ATP and some anion channel inhibitors. These findings indicate that the expression of pI(Cln) directly confers tolerance to hypotonic stress, and pI(Cln) is concluded to be an important molecule for cell-volume regulation.

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