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Inducible expression of a mutant form of MEK1 in Swiss 3T3 cells

✍ Scribed by Li-Jyun Syu; Kun-Liang Guan; Alan R. Saltiel

Publisher: John Wiley and Sons
Year: 1997
Tongue: English
Weight: 330 KB
Volume: 67
Category: Article
ISSN: 0730-2312
DOI: 10.1002/(sici)1097-4644(19971201)67:3<367::aid-jcb8>3.0.co;2-u

No coin nor oath required. For personal study only.

✦ Synopsis

We conditionally overexpressed a MEK1 mutant that contains triple mutations in the regulatory and kinase domains, and investigated its effects on the MAP kinase cascade in Swiss 3T3 cells. Expression of the mutant produced a 60% blockade in MAP kinase activity. However, only a modest blockade in DNA synthesis was observed, without any reductions in the phosphorylation of two proteins known to be substrates of MAP kinase. Moreover, the overexpression of MEK1(3A) failed to block endogenous MEK1 activation, although MEK1(3A) formed complexes with both c-Raf and B-Raf as well as p42/p44 MAPK. These results suggest that there may be multiple biochemical inputs into the MEK/MAPK pathway.

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