Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.

The proto-oncoprotein Raf is pivotal for mitogen-activated protein kinase (MAPK) signaling, and its aberrant activation has been implicated in multiple human cancers. However, the precise molecular mechanism of Raf activation, especially for B-Raf, remains unresolved. By genetic and biochemical stud...

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Main Authors: Fan Xia, Jinghong Li, Gavin W Hickey, Amy Tsurumi, Kimberly Larson, Dongdong Guo, Shian-Jang Yan, Louis Silver-Morse, Willis X Li
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2008-05-01
Series:PLoS Biology
Online Access:https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0060128&type=printable
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author Fan Xia
Jinghong Li
Gavin W Hickey
Amy Tsurumi
Kimberly Larson
Dongdong Guo
Shian-Jang Yan
Louis Silver-Morse
Willis X Li
author_facet Fan Xia
Jinghong Li
Gavin W Hickey
Amy Tsurumi
Kimberly Larson
Dongdong Guo
Shian-Jang Yan
Louis Silver-Morse
Willis X Li
author_sort Fan Xia
collection DOAJ
description The proto-oncoprotein Raf is pivotal for mitogen-activated protein kinase (MAPK) signaling, and its aberrant activation has been implicated in multiple human cancers. However, the precise molecular mechanism of Raf activation, especially for B-Raf, remains unresolved. By genetic and biochemical studies, we demonstrate that phosphorylation of tyrosine 510 is essential for activation of Drosophila Raf (Draf), which is an ortholog of mammalian B-Raf. Y510 of Draf is phosphorylated by the c-src homolog Src64B. Acidic substitution of Y510 promotes and phenylalanine substitution impairs Draf activation without affecting its enzymatic activity, suggesting that Y510 plays a purely regulatory role. We further show that Y510 regulates Draf activation by affecting the autoinhibitory interaction between the N- and C-terminal fragments of the protein. Finally, we show that Src64B is required for Draf activation in several developmental processes. Together, these results suggest a novel mechanism of Raf activation via Src-mediated tyrosine phosphorylation. Since Y510 is a conserved residue in the kinase domain of all Raf proteins, this mechanism is likely evolutionarily conserved.
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institution OA Journals
issn 1544-9173
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publishDate 2008-05-01
publisher Public Library of Science (PLoS)
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spelling doaj-art-4036ce6e4cc642a2b53bb3176313e2a32025-08-20T02:00:50ZengPublic Library of Science (PLoS)PLoS Biology1544-91731545-78852008-05-0165e12810.1371/journal.pbio.0060128Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.Fan XiaJinghong LiGavin W HickeyAmy TsurumiKimberly LarsonDongdong GuoShian-Jang YanLouis Silver-MorseWillis X LiThe proto-oncoprotein Raf is pivotal for mitogen-activated protein kinase (MAPK) signaling, and its aberrant activation has been implicated in multiple human cancers. However, the precise molecular mechanism of Raf activation, especially for B-Raf, remains unresolved. By genetic and biochemical studies, we demonstrate that phosphorylation of tyrosine 510 is essential for activation of Drosophila Raf (Draf), which is an ortholog of mammalian B-Raf. Y510 of Draf is phosphorylated by the c-src homolog Src64B. Acidic substitution of Y510 promotes and phenylalanine substitution impairs Draf activation without affecting its enzymatic activity, suggesting that Y510 plays a purely regulatory role. We further show that Y510 regulates Draf activation by affecting the autoinhibitory interaction between the N- and C-terminal fragments of the protein. Finally, we show that Src64B is required for Draf activation in several developmental processes. Together, these results suggest a novel mechanism of Raf activation via Src-mediated tyrosine phosphorylation. Since Y510 is a conserved residue in the kinase domain of all Raf proteins, this mechanism is likely evolutionarily conserved.https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0060128&type=printable
spellingShingle Fan Xia
Jinghong Li
Gavin W Hickey
Amy Tsurumi
Kimberly Larson
Dongdong Guo
Shian-Jang Yan
Louis Silver-Morse
Willis X Li
Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
PLoS Biology
title Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
title_full Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
title_fullStr Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
title_full_unstemmed Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
title_short Raf activation is regulated by tyrosine 510 phosphorylation in Drosophila.
title_sort raf activation is regulated by tyrosine 510 phosphorylation in drosophila
url https://journals.plos.org/plosbiology/article/file?id=10.1371/journal.pbio.0060128&type=printable
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