M. Genini et al., SUBTRACTIVE CLONING AND CHARACTERIZATION OF DRAL, A NOVEL LIM-DOMAIN PROTEIN DOWN-REGULATED IN RHABDOMYOSARCOMA, DNA and cell biology, 16(4), 1997, pp. 433-442
A subtractive cloning procedure was used to characterize the molecular
changes involved in transformation of normal myoblasts to rhabdomyosa
rcoma (RMS) cells. Here we describe the cloning of DRAL, a novel LIM-d
omain protein expressed in primary myoblasts but down-regulated in the
RMS cell line RD. DRAL is a LIM-only protein with five LIM domains wh
ereby one LIM domain consists only of the second half of the consensus
motif. Interestingly, down-regulation of DRAL was not confined to the
RD RMS cells, but was a phenomenon extended to other RMS cell lines o
f both embryonal and alveolar subtype, and to some breast cancer cell
lines. Analysis of the expression pattern in normal human tissues reve
aled that DRAL is expressed at high levels in the heart, suggesting an
important function in the specification of the terminally differentia
ted phenotype of heart muscle cells. Immunofluorescence studies using
an antibody directed against recombinant DRAL localized the protein pr
edominantly in the nucleus of cultured cells. On the basis of these re
sults, we conclude that down-regulation of DRAL correlates with the tu
mor phenotype of RMS cells.