Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

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

doi:10.1016/j.leukres.2006.01.001

« Previous Next »Leukemia Research
Volume 30, Issue 9 , Pages 1097-1104, September 2006

Phosphoproteomic analysis of AML cell lines identifies leukemic oncogenes

Denise K. Walters
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States
- Corresponding author. Tel.: +1 503 494 5599; fax: +1 503 494 3688.
- These authors equally contributed to this work.
Valerie L. Goss
- Cell Signaling Technology Inc., 166B Cummings Center, Beverly, MA 01915, United States
- These authors equally contributed to this work.
Eric P. Stoffregen
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States
Ting-Lei Gu
- Cell Signaling Technology Inc., 166B Cummings Center, Beverly, MA 01915, United States
Kimberly Lee
- Cell Signaling Technology Inc., 166B Cummings Center, Beverly, MA 01915, United States
Julie Nardone
- Cell Signaling Technology Inc., 166B Cummings Center, Beverly, MA 01915, United States
Laura McGreevey
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States
Michael C. Heinrich
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States
Michael W. Deininger
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States
Roberto Polakiewicz
- Cell Signaling Technology Inc., 166B Cummings Center, Beverly, MA 01915, United States
Brian J. Druker
- Department of Hematology and Oncology, Howard Hughes Medical Institute, Oregon Health and Science University Cancer Institute, 3181 Sam Jackson Park Road, and Portland VA Medical Center, Portland, OR 97239, United States

Received 5 October 2005; received in revised form 6 December 2005; accepted 3 January 2006. published online 07 March 2011.

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, Phosphopeptide, Tyrosine kinases, Leukemia