Expression cloning of oligomerization-activated genes with cell-proliferating potency by pseudotype retrovirus vector.
We developed a method of clone proliferation promoting fusion genes whose proteins were activated by protein oligomerization through the helix-loop-helix region (PNT domain) of TEL. We inserted a cDNA library downstream of the PNT domain with a retrovirus vector. The resulting retrovirus infected cytokine-dependent 32D cells and cells with cytokine-independent growth were analyzed for the inserted cDNA. We cloned 25 independent fusion genes including seven kinds of partner genes. Six of the seven were a fusion of TEL with protein tyrosine kinase, LYN, HCK, FGR, SYK, FLT3, and TYK2. A serine/threonine kinase, ARAF1, was also found to fuse with TEL. These kinase fusion proteins included kinase domains with proper reading frames. These fusions may be a useful model for clarifying the downstream signal transduction of constitutive active kinase and this expression cloning method may provide a new tool with which to study cell proliferation signalling.
Abe
Akihiro
A
Department of Hematology, Nagoya University Graduate School of Medicine, Nagoya, Japan. aakihiro@med.nagoya-u.ac.jp
Emi
Nobuhiko
N
Kanie
Tadaharu
T
Imagama
Shizuka
S
Kuno
Yoshie
Y
Takahashi
Masahide
M
Saito
Hidehiko
H
Naoe
Tomoki
T
D000818:Animals; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002472:Cell Transformation, Viral / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D019281:Dimerization; D005786:Gene Expression Regulation / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D051379:Mice; D022423:Myeloid Cells / Q000166:cytology / Q000378:metabolism*; D011108:Polymers / Q000378:metabolism; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics*; D000097764:ETS Translocation Variant 6 Protein
D004268:DNA-Binding Proteins; D011108:Polymers; D050783:Proto-Oncogene Proteins c-ets; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D000097764:ETS Translocation Variant 6 Protein