Interaction between the pRb2/p130 C-terminal domain and the N-terminal portion of cyclin D3.
An association between cyclin D3 and the C-terminal domain of pRb2/p130 was demonstrated using the yeast two-hybrid system. Further analysis restricted the epitope responsible for the binding within the 74 N-terminal amino acids of cyclin D3, independent of the LXCXE amino acid motif present in the D-type cyclin N-terminal region. In a coprecipitation assay in T98G cells, a human glioblastoma cell line, the C-terminal domain of pRb2/p130 was able to interact solely with cyclin D3, while the corresponding portion of pRb interacted with either cyclin D3 or cyclin D1. In T98G cells, endogenous cyclin D3-associated kinase activity showed a clear predisposition to phosphorylate preferentially the C-terminal domain of pRb2/p130, rather than that of pRb. This propensity was also confirmed in LAN-5 human neuroblastoma cells, where phosphorylation of the pRb2/p130 C-terminal domain and expression of cyclin D3 also decreased remarkably in the late neural differentiation stages.
Bonetto
F
F
Laboratory for Cell Metabolism and Pharmacokinetics, Center for Experimental Research, Regina Elena Cancer Institute, 00158 Rome, Italy.
Fanciulli
M
M
Battista
T
T
De Luca
A
A
Russo
P
P
Bruno
T
T
De Angelis
R
R
Di Padova
M
M
Giordano
A
A
Felsani
A
A
Paggi
M G
MG
D017500:Adenovirus E1A Proteins / Q000235:genetics / Q000378:metabolism; D020816:Amino Acid Motifs / Q000235:genetics; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015153:Blotting, Western; D019938:Cyclin D1 / Q000378:metabolism; D056743:Cyclin D3; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016213:Cyclins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D011506:Proteins; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D050717:Retinoblastoma-Like Protein p130; D020798:Two-Hybrid System Techniques
D017500:Adenovirus E1A Proteins; D000906:Antibodies; C534696:CCND3 protein, human; C534697:Ccnd3 protein, mouse; D056743:Cyclin D3; D016213:Cyclins; D010455:Peptides; D010750:Phosphoproteins; D011506:Proteins; C493837:RBL2 protein, human; C493838:Rbl2 protein, mouse; D011993:Recombinant Fusion Proteins; D050717:Retinoblastoma-Like Protein p130; D019938:Cyclin D1; D018844:Cyclin-Dependent Kinases