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TRA1, a novel mRNA highly expressed in leukemogenic mouse monocytic
sublines but not in nonleukemogenic sublines
T Kasukabe, J Okabe-Kado and Y Honma
Department of Chemotherapy, Saitama Cancer Center Research Institute, Ina,
Japan.
Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse
monocytic and immortalized Mm-1 cells derived from spontaneously
differentiated, mouse myeloblastic M1 cells. Although these subline cells
retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are
highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas
Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better
understand the molecular mechanisms of these levels of leukemogenicity, we
investigated putative leukemogenesis- associated genes or oncogenes
involved in the maintenance of growth, especially in vivo, by means of
differential mRNA display. We isolated a fragment clone (15T01) from Mm-P
cells. The mRNA probed with 15T01 was expressed at high levels in
leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2
cells. The gene corresponding to 15T01, named TRA1, was isolated from an
Mm-P cDNA library. The longest open reading frame of the TRA1 clone
predicts a peptide containing 204 amino acids with a calculated molecular
weight of 23,049 D. The predicted TRA1 protein is cysteine-rich and
contains multiple cysteine doublets. A putative normal counterpart gene,
named NOR1, was also isolated from a normal mouse kidney cDNA library and
sequenced. NOR1 cDNA predicts a peptide containing 234 amino acids. The
sequence of 201 amino acids from the C-terminal NOR1 was completely
identical to that of TRA1, whereas the remaining N-terminal amino acids (33
amino acids) were longer than that (3 amino acids) of TRA1 and the
N-terminus of NOR1 protein contained proline-rich sequence. A similarity
search against current nucleotide and protein sequence databases indicated
that the NOR1/TRA1 gene(s) is conserved in a wide range of eukaryotes,
because apparently homologous genes were identified in Caenorhabditis
elegans and Saccharomyces cerevisiae genomes. Northern blotting using
TRA1-specific and NOR1-specific probes indicated that TRA1 mRNA is
exclusively expressed in leukemogenic but not in nonleukemogenic Mm
sublines and normal tissues and also indicated that NOR1 mRNA is expressed
in normal tissues, especially in kidney, lung, liver, and bone marrow cells
but not in any Mm sublines. After leukemogenic Mm-P cells were induced to
differentiate into normal macrophages by sodium butyrate, the normal
counterpart, NOR1, was expressed, whereas the TRA1 level decreased.
Furthermore, transfection of TRA1 converted nonleukemogenic Mm-S1 cells
into leukemogenic cells. These results indicate that the TRA1 gene is
associated at least in part with the leukemogenesis of monocytic Mm
sublines.
Volume 89,
Issue 8,
pp. 2975-2985,
04/15/1997
Copyright © 1997 by The American Society of Hematology

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