Expression of human tyrosine kinase, Lck, in yeast Saccharomyces cerevisiae: growth suppression and strategy for inhibitor screening.
We report the successful expression and detection of a phosphorylated form of human T cell tyrosine kinase, Lck, in Saccharomyes cerevisiae, which leads to growth suppression of the yeast cells. Expression of an inactive Lck mutant resulted in no phosphorylation and no growth suppression, indicating that cell growth inhibition by Lck is due to the activity of the kinase, consistent with the observed tyrosine-phosphorylation of the Lck and yeast host cell proteins. The addition of a known inhibitor of Lck to the cell culture resulted in recovery of cell growth expressing the active Lck, suggesting that the growth inhibition by lck gene expression can be used to screen inhibitors for the gene product. We have extended such approach to Tob, another potential therapeutic target.
Koyama
Masashi
M
Epoch Medical International Inc. EMI, KI, Osaka, Japan.
Saito
Shoko
S
Nakagawa
Rumiko
R
Katsuyama
Iwao
I
Hatanaka
Masakazu
M
Yamamoto
Tadashi
T
Arakawa
Tsutomu
T
Tokunag
Masao
M
D004791:Enzyme Inhibitors / Q000378:metabolism*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck) / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010766:Phosphorylation; D011743:Pyrimidines / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
C412373:AG 1879; D004791:Enzyme Inhibitors; D047908:Intracellular Signaling Peptides and Proteins; D011743:Pyrimidines; C098669:TOB1 protein, human; D025521:Tumor Suppressor Proteins; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck)