Mammalian Ras interacts directly with the serine/threonine kinase Raf
AB Vojtek, SM Hollenberg, JA Cooper - Cell, 1993 - cell.com
AB Vojtek, SM Hollenberg, JA Cooper
Cell, 1993•cell.comWe have identified proteins that interact with H-Ras using a two hybrid system screen of a
mouse cDNA library. Approximately 50% of the clones identified encoded portionsof the c-
Raf and A-Rat serinelthreonine kinases. Overlaps among these clones define a conserved
81 residue region of the N-terminus of Raf as the Ras interaction region. We show that Raf
interacts with wild-type and activated Ras, but not with an effector domain mutant of Ras or
with a dominantinterfering Ras mutant. Using purified bacterially expressed fusion proteins …
mouse cDNA library. Approximately 50% of the clones identified encoded portionsof the c-
Raf and A-Rat serinelthreonine kinases. Overlaps among these clones define a conserved
81 residue region of the N-terminus of Raf as the Ras interaction region. We show that Raf
interacts with wild-type and activated Ras, but not with an effector domain mutant of Ras or
with a dominantinterfering Ras mutant. Using purified bacterially expressed fusion proteins …
Summary
We have identified proteins that interact with H-Ras using a two hybrid system screen of a mouse cDNA library. Approximately 50% of the clones identified encoded portionsof the c-Raf and A-Rat serinelthreonine kinases. Overlaps among these clones define a conserved 81 residue region of the N-terminus of Raf as the Ras interaction region. We show that Raf interacts with wild-type and activated Ras, but not with an effector domain mutant of Ras or with a dominantinterfering Ras mutant. Using purified bacterially expressed fusion proteins, we show, furthermore, that Ras and the N-terminal region of Raf associate directly in vitro and that this interaction Is dependent on GTP bound to Ras.
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