Comparison of protein tyrosine phosphorylation and morphological changes induced by IL-2 and IL-3.
We constructed cell lines which can proliferate in response to IL-2 or IL-3 by introducing a wild-type and mutant forms of cDNAs encoding the human IL-2R p75 chain into an IL-3 dependent hematopoietic cell line which expresses the p55 chain of the IL-2R. We compared early events that were induced in these cells by IL-2 and IL-3. Analysis of protein tyrosine phosphorylation showed that two common protein bands, pp95 and pp90, were phosphorylated by stimulation of either IL-2 or IL-3, suggesting the possible sharing of part of a signal transduction pathway between IL-2R and IL-3R. Comparison of protein tyrosine phosphorylation profiles induced by IL-2 and IL-3 among a variety of cell lines revealed that the pp90 band is the common tyrosine phosphorylation substrate in the cell lines examined, although the general tyrosine phosphorylation pattern differed in each cell line. Mutant p75 molecules incapable of inducing tyrosine phosphorylation could bind and internalize IL-2, but could not support cell growth. We also found that swift changes of cytoskeletal protein organization are one of the early events caused by signal transduction through either IL-2R and IL-3R. Reorganization of cytoskeletal proteins seems to be associated with protein phosphorylation, as a significant portion of pp90 was found in a detergent-soluble fraction in IL-2 or IL-3 treated cells.
Sabe
H
H
Center for Molecular Biology and Genetics, Kyoto University Faculty of Medicine, Japan.
Kuno
J
J
Koromilas
A
A
Saito
Y
Y
Kinashi
T
T
Ueda
M
M
Takamatsu
T
T
Hamaguchi
M
M
Kawakami
T
T
Honjo
T
T
D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000235:genetics; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology*; D007377:Interleukin-3 / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009154:Mutation; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D015375:Receptors, Interleukin-2 / Q000235:genetics / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
D007376:Interleukin-2; D007377:Interleukin-3; D011506:Proteins; D015375:Receptors, Interleukin-2; D014443:Tyrosine; D004247:DNA