Phosphorylation site substrate specificity determinants for the PIM-1 protooncogene-encoded protein kinase

Palaty, Chrystal K.; Clark‐Lewis, Ian; Leung, Dan; Pelech, Steven

doi:10.1139/o97-026

Cited by 48 publications

(23 citation statements)

References 43 publications

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“…This is consistent with the general lack of selectivity at the downstream positions and with previous observations that deletion of these residues in peptide substrates does not affect Pim-1 catalytic efficiency (33). The Pim-1 structure provides a potential rationale for these observations.…”

Section: Structures Of Pim-1 In Complex With a Consensus Peptide And supporting

confidence: 91%

“…Sequence-Based on analysis of individual peptide substrates, Pim-1 has been described as having selectivity for basic residues at the Ϫ5 through Ϫ2 positions relative to the phosphorylation site and for smaller residues at the ϩ1 position (32,33). Recently we used a peptide library approach to systematically evaluate the contribution of all amino acid residues at each of nine positions surrounding a fixed phosphoacceptor site to phosphorylation by Pim-2 (34).…”

Section: The Three Pim Kinases Share a Common Phosphorylation Consensusmentioning

confidence: 99%

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Structure and Substrate Specificity of the Pim-1 Kinase

Bullock

Debreczeni

Amos

et al. 2005

Journal of Biological Chemistry

176

199

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The Pim kinases are a family of three vertebrate protein serine/ threonine kinases (Pim-1, -2, and -3) belonging to the CAMK (calmodulin-dependent protein kinase-related) group. Pim kinases are emerging as important mediators of cytokine signaling pathways in hematopoietic cells, and they contribute to the progression of certain leukemias and solid tumors. A number of cytoplasmic and nuclear proteins are phosphorylated by Pim kinases and may act as their effectors in normal physiology and in disease. Recent crystallographic studies of Pim-1 have identified unique structural features but have not provided insight into how the kinase recognizes its target substrates. Here, we have conducted peptide library screens to exhaustively determine the sequence specificity of active site-mediated phosphorylation by Pim-1 and Pim-3. We have identified the major site of Pim-1 autophosphorylation and find surprisingly that it maps to a novel site that diverges from its consensus phosphorylation motif. We have solved the crystal structure of Pim-1 bound to a high affinity peptide substrate in complexes with either the ATP analog AMP-PNP or the bisindolylmaleimide kinase inhibitor 2-[1-(3-dimethylaminopropyl)-1H-indol-3-yl]-3-(1H-indol-3-yl)maleimide HCl. These structures reveal an unanticipated mode of recognition for basic residues upstream of the phosphorylation site, distinct from that of other kinases with similar substrate specificity. The structures provide a rationale for the unusually high affinity of Pim kinases for peptide substrates and suggest a general mode for substrate binding to members of the CAMK group.Pim-1 was first described with c-myc as a frequent proviral insertion site in Moloney murine leukemia virus-induced T-cell lymphomas (1, 2). Activation of the two oncogenes is strongly cooperative, such that double-transgenic E-myc E-pim1 mice exhibit pre-B-cell leukemia in utero (3). Likewise, activating proviral insertion in the pim2 and pim3 genes can also contribute to leukemia in the mouse (4, 5). Pim-1 overexpression is observed in a range of human lymphomas and acute leukemias (6), whereas Pim-2 overexpression is associated with chronic lymphocytic leukemia and non-Hodgkin lymphoma (7). Pim-1 is also a prognostic marker in prostate cancer (8,9), and Pim-3 shows aberrant expression in hepatocellular carcinoma (10). Additionally, pim1 somatic hypermutations (11) and chromosomal translocations have been identified in non-Hodgkin lymphoma (12). In light of their oncogenic potential, the Pim kinase family is emerging as an important new target for drug discovery efforts (13-15).Physiologically, Pim kinases appear to contribute to the proliferation and survival of leukocytes. Pim-1 and Pim-2 are expressed at low levels in many tissues but are strongly induced in leukocytes by the JAK/ STAT 2 pathway in response to cytokines, including interleukins 2, 3, and 7, granulocyte-macrophage colony-stimulating factor, ␣-and ␥-interferon, and erythropoietin (16 -18). Pim kinase regulation appears to be largely at the le...

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Section: Structures Of Pim-1 In Complex With a Consensus Peptide And supporting

confidence: 91%

Section: The Three Pim Kinases Share a Common Phosphorylation Consensusmentioning

confidence: 99%

Structure and Substrate Specificity of the Pim-1 Kinase

Bullock

Debreczeni

Amos

et al. 2005

Journal of Biological Chemistry

176

199

View full text Add to dashboard Cite

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“…[20][21][22][23] Functional interactions between the PIM family and MYC have been most well characterized in genetic rodent models of lymphomagenesis. 17 Employing biochemical and overexpression methodologies, PIM kinases have been shown to regulate MYC protein stability through direct phosphorylation of MYC S329.…”

Section: Introductionmentioning

confidence: 99%

PIM inhibitors target CD25-positive AML cells through concomitant suppression of STAT5 activation and degradation of MYC oncogene

Guo

Wang

Zhang

et al. 2014

Blood

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Key Points• CD25 is a predictive biomarker for sensitivity to PIM inhibitors in AML cells.• PIM inhibitors may prolong overall/relapse-free survival through attenuating STAT5 activation and destabilizing MYC in CD25 1 AML cells.Postchemotherapy relapse presents a major unmet medical need in acute myeloid leukemia (AML), where treatment options are limited. CD25 is a leukemic stem cell marker and a conspicuous prognostic marker for overall/relapse-free survival in AML. Rare occurrence of genetic alterations among PIM family members imposes a substantial hurdle in formulating a compelling patient stratification strategy for the clinical development of selective PIM inhibitors in cancer. Here we show that CD25, a bona fide STAT5 regulated gene, is a mechanistically relevant predictive biomarker for sensitivity to PIM kinase inhibitors. Alone or in combination with tyrosine kinase inhibitors, PIM inhibitors can suppress STAT5 activation and significantly shorten the half-life of MYC to achieve substantial growth inhibition of high CD25-expressing AML cells. Our results highlight the importance of STAT5 and MYC in rendering cancer cells sensitive to PIM inhibitors. Because the presence of a CD25-positive subpopulation in leukemic blasts correlates with poor overall or relapse-free survival, our data suggest that a combination of PIM inhibitors with chemotherapy and tyrosine kinase inhibitors could improve long-term therapeutic outcomes in

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“…Hsp90 is coordinately regulated with PIM-1 and is responsible for the stabilization and function of PIM-1 (24,25). PIM-1 is able to phosphorylate itself (26,27) through its recently identified novel autophosphorylation site that diverges from its consensus phosphorylation motif (28). Several substrates of PIM-1 have been identified, including p21Cip1/WAF1 (29,30), Cdc25A (31), PTPU2 (32), NuMA (33), C-TAK1 (34), and Cdc25C (35), indicating PIM-1 is involved in the cell proliferation at both G 1 /S and G 2 /M transition.…”

Section: Introductionmentioning

confidence: 99%

PIM-1–specific mAb suppresses human and mouse tumor growth by decreasing PIM-1 levels, reducing Akt phosphorylation, and activating apoptosis

Hu¹,

Li²,

Vandervalk³

et al. 2009

J. Clin. Invest.

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Provirus integration site for Moloney murine leukemia virus (PIM1) is a proto-oncogene that encodes a serine/ threonine kinase with multiple cellular functions. Overexpression of PIM-1 plays a critical role in progression of prostatic and hematopoietic malignancies. Here we describe the generation of a mAb specific for GST-PIM-1, which reacted strongly with most human and mouse cancer tissues and cell lines of prostate, breast, and colon origin but only weakly (if at all) with normal tissues. The mAb binds to PIM-1 in the cytosol and nucleus as well as to PIM-1 on the surface of human and murine cancer cells. Treatment of human and mouse prostate cancer cell lines with the PIM-1-specific mAb resulted in disruption of PIM-1/Hsp90 complexes, decreased PIM-1 and Hsp90 levels, reduced Akt phosphorylation at Ser473, reduced phosphorylation of Bad at Ser112 and Ser136, and increased cleavage of caspase-9, an indicator of activation of the mitochondrial cell death pathway. The mAb induced cancer cell apoptosis and synergistically enhanced antitumor activity when used in combination with cisplatin and epirubicin. In tumor models, the PIM-1-specific mAb substantially inhibited growth of the human prostate cancer cell line DU145 in SCID mice and the mouse prostate cancer cell TRAMP-C1 in C57BL/6 mice. These findings are important because they provide what we believe to be the first in vivo evidence that treatment of prostate cancer may be possible by targeting PIM-1 using an Ab-based therapy.

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Phosphorylation site substrate specificity determinants for the PIM-1 protooncogene-encoded protein kinase

Cited by 48 publications

References 43 publications

Structure and Substrate Specificity of the Pim-1 Kinase

Structure and Substrate Specificity of the Pim-1 Kinase

PIM inhibitors target CD25-positive AML cells through concomitant suppression of STAT5 activation and degradation of MYC oncogene

PIM-1–specific mAb suppresses human and mouse tumor growth by decreasing PIM-1 levels, reducing Akt phosphorylation, and activating apoptosis

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