Identification of RB18A, a 205 kDa new p53 regulatory protein which shares antigenic and functional properties with p53.
Immunological screening with the anti-p53 moAb, PAb1801 of a cDNA expression library, prepared from human B lymphoma cells, led us to identify a new human 205 kDa protein called RB18A for 'Recognized By PAb1801 moAntibody'. Immunoblotting or immunoprecipitation of fusion protein or in vitro translated protein, respectively, demonstrated that RB18A protein was recognized by several anti-p53 moAb reacting with the N or C-terminal domains of p53. Full length sequence of RB18A cDNA and computer analysis demonstrated that despite common antigenic determinants between RB18A and p53 proteins, nucleotide and deduced protein sequences did not reveal any significant homologies. RB18A mRNA was detected in all tissues tested except in kidney. In addition, RB18A protein shared identical functions with p53 protein: binding to DNA or to p53 and self-oligomerization. Furthermore, RB18A regulated p53 specific binding on his DNA consensus binding site. These functions were associated to the C-terminal domain of RB18A protein and more specifically to the PAb421 binding site present in this domain. The activation by RB18A of p53 binding on DNA was induced through an unstable interaction between both proteins. Altogether, our data demonstrated that RB18A protein shares antigenic and functional properties with p53 and regulated p53 functions.
Drané
P
P
Immunochimie des Régulations Cellulaires et des Interactions Virales, Inserm U.354, Centre Inserm, Hôpital Saint-Antoine, Paris, France.
Barel
M
M
Balbo
M
M
Frade
R
R
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification*; D018076:DNA, Complementary / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D016393:Lymphoma, B-Cell / Q000235:genetics*; D056913:Mediator Complex Subunit 1; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000302:isolation & purification*; D014407:Tumor Cells, Cultured / Q000276:immunology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification* / Q000502:physiology
D002352:Carrier Proteins; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C538855:MED1 protein, human; D056913:Mediator Complex Subunit 1; D012333:RNA, Messenger; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53