Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

Denise K. Walters; Valerie L. Goss; Eric P. Stoffregen; Ting Lei Gu; Kimberly Lee; Julie Nardone; Laura McGreevey; Michael C. Heinrich; Michael W. Deininger; Roberto Polakiewicz; Brian J. Druker

doi:10.1016/j.leukres.2006.01.001

Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

Denise K. Walters, Valerie L. Goss, Eric P. Stoffregen, Ting Lei Gu, Kimberly Lee, Julie Nardone, Laura McGreevey, Michael C. Heinrich, Michael W. Deininger, Roberto Polakiewicz, Brian J. Druker

Knight Cancer Institute

Research output: Contribution to journal › Article › peer-review

52 Scopus citations

Abstract

STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.

Original language	English (US)
Pages (from-to)	1097-1104
Number of pages	8
Journal	Leukemia Research
Volume	30
Issue number	9
DOIs	https://doi.org/10.1016/j.leukres.2006.01.001
State	Published - Sep 2006

Keywords

AML
Leukemia
Phosphopeptide
Tyrosine kinases

ASJC Scopus subject areas

Hematology
Oncology
Cancer Research

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10.1016/j.leukres.2006.01.001

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@article{7dff62f7c61c429abdef3f1340126104,

title = "Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes",

abstract = "STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.",

keywords = "AML, Leukemia, Phosphopeptide, Tyrosine kinases",

author = "Walters, {Denise K.} and Goss, {Valerie L.} and Stoffregen, {Eric P.} and Gu, {Ting Lei} and Kimberly Lee and Julie Nardone and Laura McGreevey and Heinrich, {Michael C.} and Deininger, {Michael W.} and Roberto Polakiewicz and Druker, {Brian J.}",

note = "Funding Information: The Howard Hughes Medical Institute, TJ Martell Foundation and Leukemia & Lymphoma Society, a grant from the Doris Duke Charitable Foundation and funding from a Veterans Affairs Merit Review Grant supported this work. This study was also funded by cell signaling for the authors at cell signaling. The initials of these authors is as follows: VG, TLG, KL, JN and RP. ",

year = "2006",

month = sep,

doi = "10.1016/j.leukres.2006.01.001",

language = "English (US)",

volume = "30",

pages = "1097--1104",

journal = "Leukemia Research",

issn = "0145-2126",

publisher = "Elsevier Limited",

number = "9",

}

TY - JOUR

T1 - Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

AU - Walters, Denise K.

AU - Goss, Valerie L.

AU - Stoffregen, Eric P.

AU - Gu, Ting Lei

AU - Lee, Kimberly

AU - Nardone, Julie

AU - McGreevey, Laura

AU - Heinrich, Michael C.

AU - Deininger, Michael W.

AU - Polakiewicz, Roberto

AU - Druker, Brian J.

N1 - Funding Information: The Howard Hughes Medical Institute, TJ Martell Foundation and Leukemia & Lymphoma Society, a grant from the Doris Duke Charitable Foundation and funding from a Veterans Affairs Merit Review Grant supported this work. This study was also funded by cell signaling for the authors at cell signaling. The initials of these authors is as follows: VG, TLG, KL, JN and RP.

PY - 2006/9

Y1 - 2006/9

N2 - STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.

AB - STAT5 is constitutively phosphorylated in leukemic cells in approximately 70% of acute myeloid leukemia (AML) patients. To identify kinase candidates potentially responsible for STAT5 phosphorylation, we used liquid chromatography-tandem mass spectrometry (LC-MS/MS) mass spectrometry to detect phosphoproteins in AML cell lines. We established TEL-ARG and BCR-ABL fusion proteins as the mechanism underlying STAT5 phosphorylation in HT-93 and KBM-3 cells, respectively. In addition, we identified a JAK2 pseudokinase domain mutation in HEL cells and using siRNA downregulation, established JAK2 as the kinase responsible for phosphorylating STAT5. This study illustrates the benefit of LC-MS/MS mass spectrometry and siRNA for the identification of novel targets and mutations.

KW - AML

KW - Leukemia

KW - Phosphopeptide

KW - Tyrosine kinases

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U2 - 10.1016/j.leukres.2006.01.001

DO - 10.1016/j.leukres.2006.01.001

M3 - Article

C2 - 16464493

AN - SCOPUS:33745717815

SN - 0145-2126

VL - 30

SP - 1097

EP - 1104

JO - Leukemia Research

JF - Leukemia Research

IS - 9

ER -

Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

Abstract

Keywords

ASJC Scopus subject areas

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