Recently, we demonstrated that the DAN gene product contains a growth-
and/or a tumor-suppressive activity in vitro, In the present work, us
ing a yeast two-hybrid system, we searched for cellular proteins that
can associate with the DAN gene product, A cDNA clone, termed DA41, wa
s initially isolated from an adult rat lung cDNA library, The DA41 gen
e was expressed in all adult tissues examined, however, the levels of
expression varied significantly among the different tissues. Like the
DAN gene product, the DA41 protein was similarly restricted to the cyt
oplasm. Sequence analysis revealed that DA41 cDNA is 2,167 nucleotides
in length and contains a single open reading frame (ORF) of 582 amino
acids (61,945 daltons), A homology search revealed that the DA41 gene
product shares no structural similarity with those filed in the data
base, In a synchronous 3Y1 cell culture, DA41 mRNA was expressed at a
low level in quiescent cells; however, its level was significantly inc
reased between the G(1) and S phases of the cell cycle, On the other h
and, the expression level of DAN mRNA did not change throughout the ce
ll cycle progression, These results suggest that the DAN-DA41 complex
might play a crucial role in the regulation of the cell cycle progress
ion.