| 15721285 |
Maeda M, Ishida A, Ni L, Kobayashi A: Isolation of CHO-K1 clones defective in cAMP-dependent proteolysis, as determined by the stability of exogenously expressed GATA-6. Biochem Biophys Res Commun. 2005 Apr 1;329(1):140-6.
Degradation of the GATA-6 (Delta50) protein expressed in a CHO-K1 clone (tc1-17a) is stimulated in the presence of dbcAMP through proteasome without new protein synthesis [FEBS Lett. 408 (1997) 301], whereas the intrinsic GC-box-binding protein was stable. To examine the cellular mechanism responsible for this specific degradation of GATA-6 (Delta50), we initially introduced the blasticidin-S deaminase gene carrying a promoter with GATA motifs that are recognized by GATA-6. The resulting cell line (tc2G2) grew in the presence of blasticidin S. However, the presence of both blasticidin S and dbcAMP was lethal due to degradation of GATA-6. Cells resistant to such lethality were isolated by chemical mutagenesis. The GATA-6 (Delta50) in these resistant cells was stable in the presence of dbcAMP in contrast to that in the parent tc2G2 cells, as determined by gel-mobility shift analysis and Western blotting. These clones could be beneficial for identification and characterization of the components participating in the signaling pathway for both protein degradation and cAMP-dependent biological processes. |
165(2,2,2,5) |