Mutations in the SAM domain of the ETV6-NTRK3 chimeric tyrosine kinase block polymerization and transformation activity

被引:35
作者
Tognon, CE
Mackereth, CD
Somasiri, AM
McIntosh, LP
Sorensen, PHB
机构
[1] BC Res Inst Childrens & Womens Hlth, Dept Pathol, Vancouver, BC V5Z 4H4, Canada
[2] BC Res Inst Childrens & Womens Hlth, Dept Pediat, Vancouver, BC V5Z 4H4, Canada
[3] Univ British Columbia, Dept Biochem & Mol Biol, Vancouver, BC V6T 1Z3, Canada
[4] Univ British Columbia, Dept Chem & Biotechnol Lab, Vancouver, BC V6T 1Z3, Canada
[5] Univ British Columbia, Dept Anat, Vancouver, BC V6H 3V5, Canada
关键词
D O I
10.1128/MCB.24.11.4636-4650.2004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The 12p13 ETV6 (TEL) gene is frequently targeted by chromosomal translocations in human malignancies, resulting in the formation of oncogenic ETV6 gene fusions. Many of the known partner genes encode protein tyrosine kinases (PTKs), generating fusion proteins that function as chimeric PTKs. ETV6-NTRK3 (EN), comprised of the ETV6 SAM domain fused to the NTRK3 PTK, is unique among ETV6 chimeric oncoproteins, as it is expressed in cancers of multiple lineages. We initially hypothesized that, similar to other ETV6-PTK chimeras, SAM-mediated dimerization of EN leads to constitutive activation of the PTK and downstream signaling cascades. However, when the EN SAM domain was replaced with an inducible FK506 binding protein (FKBP) dimerization system, resulting FKBP-NTRK3 chimeras failed to transform NIH 3T3 cells even though PTK activation was preserved. It was recently shown that the ETV6 SAM domain has two potential interacting surfaces, raising the possibility that this domain can mediate protein polymerization. We therefore mutated each EN SAM binding interface in a manner shown previously to abolish self-association of wild-type ETV6. Each mutation completely blocked the ability of EN to polymerize, to activate its PTK, and to transform NIH 3T3 cells. Furthermore, EN itself formed large polymeric structures within cells while mutant EN proteins were present only as monomers. Finally, we observed a dominant negative effect on the transformation of isolated SAM domains coexpressed in EN-transformed cells. Taken together, our results suggest that higher-order polymerization may be a critical requirement for the transformation activity of EN and possibly other ETV6-PTK fusion proteins.
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收藏
页码:4636 / 4650
页数:15
相关论文
共 53 条
[1]   Targeting of the N-terminal coiled coil oligomerization interface of BCR interferes with the transformation potential of BCR-ABL and increases sensitivity to STI571 [J].
Beissert, T ;
Puccetti, E ;
Bianchini, A ;
Güller, S ;
Boehrer, S ;
Hoelzer, D ;
Ottmann, OG ;
Nervi, C ;
Ruthardt, M .
BLOOD, 2003, 102 (08) :2985-2993
[2]   Oncogenic kinase signalling [J].
Blume-Jensen, P ;
Hunter, T .
NATURE, 2001, 411 (6835) :355-365
[3]   Fusion genes in leukemia: an emerging network [J].
Bohlander, SK .
CYTOGENETICS AND CELL GENETICS, 2000, 91 (1-4) :52-56
[4]   The tyrosine kinase Abl-related gene ARG is fused to ETV6 in an AML-M4Eo patient with a t(1;12)(q25;p13):: Molecular cloning of both reciprocal transcripts [J].
Cazzaniga, G ;
Tosi, S ;
Aloisi, A ;
Giudici, G ;
Daniotti, M ;
Pioltelli, P ;
Kearney, L ;
Biondi, A .
BLOOD, 1999, 94 (12) :4370-4373
[5]   Solution structure of a conserved C-terminal domain of p73 with structural homology to the SAM domain [J].
Chi, SW ;
Ayed, A ;
Arrowsmith, CH .
EMBO JOURNAL, 1999, 18 (16) :4438-4445
[6]   Fusion of ETV6 to neurotrophin-3 receptor TRKC in acute myeloid leukemia with t(12;15)(p13;q25) [J].
Eguchi, M ;
Eguchi-Ishimae, M ;
Tojo, A ;
Morishita, K ;
Suzuki, K ;
Sato, Y ;
Kudoh, S ;
Tanaka, K ;
Setoyama, M ;
Nagamura, F ;
Asano, S ;
Kamada, N .
BLOOD, 1999, 93 (04) :1355-1363
[7]   FUSION OF PDGF RECEPTOR-BETA TO A NOVEL ETS-LIKE GENE, TEL, IN CHRONIC MYELOMONOCYTIC LEUKEMIA WITH T(512) CHROMOSOMAL TRANSLOCATION [J].
GOLUB, TR ;
BARKER, GF ;
LOVETT, M ;
GILLILAND, DG .
CELL, 1994, 77 (02) :307-316
[8]  
Golub TR, 1996, MOL CELL BIOL, V16, P4107
[9]   FUSION OF THE TEL GENE ON 12P13 TO THE AML1 GENE ON 21Q22 IN ACUTE LYMPHOBLASTIC-LEUKEMIA [J].
GOLUB, TR ;
BARKER, GF ;
BOHLANDER, SK ;
HIEBERT, SW ;
WARD, DC ;
BRAYWARD, P ;
MORGAN, E ;
RAIMONDI, SC ;
ROWLEY, JD ;
GILLILAND, DG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (11) :4917-4921
[10]   RNA recognition via the SAM domain of Smaug [J].
Green, JB ;
Gardner, CD ;
Wharton, RP ;
Aggarwal, AK .
MOLECULAR CELL, 2003, 11 (06) :1537-1548