Complexity of translationally controlled transcription factor Sp3 isoform expression.
Sp3 is a ubiquitous transcription factor closely related to Sp1. Both proteins contain a highly conserved DNA-binding domain close to the C terminus and two glutamine-rich domains in the N-terminal moiety. Immunoblot analyses of Sp3 reveal a striking complex protein pattern of up to eight distinct species. This pattern is not observed in Sp3-deficient cell lines showing that all signals reflect Sp3 antigen. In this study, we have unraveled the complexity of Sp3 expression. We show that four isoforms of Sp3 that retain different parts of the N terminus are expressed in vivo. The four isoforms derive from alternative translational start sites at positions 1, 37, 856, and 907. An upstream open reading frame located at position -47 to -18 regulates expression of the two long isoforms. Unlike Sp1, none of the Sp3 isoforms is glycosylated. However, all four isoforms become SUMO-modified in vivo and in vitro specifically and exclusively at lysine residue 551. The transcriptional activity of the two long isoforms strongly depends on the promoter settings, whereas the small isoforms appear to be inactive. The transcriptional activity of all the Sp3 isoforms is regulated by SUMO modification. Our results demonstrate that Sp3 has many unique features and is not simply a functional equivalent of Sp1.
Sapetschnig
Alexandra
A
Institute of Molecular Biology and Tumor Research, Philipps-University, Emil-Mannkopff-Strasse 2, D-35033 Marburg, Germany.
Koch
Franziska
F
Rischitor
Grigore
G
Mennenga
Trientje
T
Suske
Guntram
G
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D018387:Codon, Initiator; D004268:DNA-Binding Proteins / Q000096:biosynthesis*; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D014176:Protein Biosynthesis / Q000502:physiology*; D020033:Protein Isoforms / Q000096:biosynthesis; D011499:Protein Processing, Post-Translational; D051705:Sp3 Transcription Factor; D014157:Transcription Factors / Q000096:biosynthesis*; D014162:Transfection; D025801:Ubiquitin / Q000378:metabolism
D018387:Codon, Initiator; D004268:DNA-Binding Proteins; D020033:Protein Isoforms; C497121:SP3 protein, human; C497122:Sp3 protein, mouse; D014157:Transcription Factors; D025801:Ubiquitin; D051705:Sp3 Transcription Factor