The block of adipocyte differentiation by a C-terminally truncated, but not by full-length, simian virus 40 large tumor antigen is dependent on an intact retinoblastoma susceptibility protein family binding domain.
Simian virus 40 (SV40) can promote cell transformation and suppress differentiation. It does this partly by targeting tumor suppressors such as p53 and members of the retinoblastoma susceptibility protein (Rb) family. This work concentrates on mechanisms by which SV40 large tumor antigen (SVLT) suppresses adipocyte differentiation. We created cell lines derived from murine 3T3-L1 preadipocytes expressing different versions of SV40 early-region sequences. SVLT-expressing cells failed to exhibit adipocyte morphology, to induce glycerophosphate dehydrogenase activity, and to induce differentiation-dependent mRNA for adipocyte P2. SVLT alone was sufficient, in the absence of SV40 small tumor antigen, to inhibit differentiation. A truncated SVLT containing only the N-terminal 121 amino acids (SVLT1-121) blocked differentiation, thus mapping at least one differentiation blocking function to the N-terminal region. K1 (Glu-107-->Lys) point mutants of SVLT, which are unable to bind to the Rb protein family or induce neoplastic transformation, are defective for blocking differentiation in the case of SVLT1-121 but retain the ability to block differentiation in the case of full-length SVLT. This finding demonstrates that Rb family proteins are important in regulating adipocyte differentiation but that other functions of full-length SVLT can block adipocyte differentiation independently of RB family binding and transformation.
Higgins
C
C
Department of Physiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Chatterjee
S
S
Cherington
V
V
D016475:3T3 Cells; D017667:Adipocytes / Q000166:cytology*; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000737:chemistry / Q000502:physiology*; D001483:Base Sequence; D001665:Binding Sites; D002448:Cell Adhesion; D002452:Cell Count; D002454:Cell Differentiation; D004279:DNA, Viral; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010446:Peptide Fragments / Q000502:physiology; D016160:Retinoblastoma Protein / Q000378:metabolism*; D013539:Simian virus 40 / Q000502:physiology*
D000952:Antigens, Polyomavirus Transforming; D004279:DNA, Viral; D010446:Peptide Fragments; D016160:Retinoblastoma Protein