Sp1 phosphorylation is involved in myelin basic protein gene transcription.
Myelin basic protein (MBP), which helps form compact myelin sheets, is a major protein expressed during oligodendrocyte (OL) differentiation. Myelin basic protein expression is regulated mainly at the transcriptional level. Previous studies showed that the transcription factor Sp1 can activate the MBP promoter. Data from the laboratory also indicate that Sp1 is expressed highly in both growing and differentiated cells. Because this is true, we wanted to understand how Sp1 activity is regulated such that it increases MBP gene transcription only in differentiating cells. Phosphorylation is one major way to regulate transcription factor activity. Our results show that there is more Sp1 binding to the MBP promoter in differentiating OLs. Sp1 is also more phosphorylated in differentiating OLs than in precursor cells. Using inhibitors of different pathways, we found that the protein kinase C (PKC) modulator phorbol 12-myristate 13-acetate (PMA) can increase Sp1 phosphorylation when the cells are treated for 1 hr and can decrease Sp1 phosphorylation with long treatment (12 hr). The increased phosphorylation of Sp1 induced by PMA in short treatments could be abolished by the extracellular signal-regulated kinases (ERK) pathway inhibitor PD98059. This indicates that PKC phosphorylates Sp1 through the ERK pathway. Mutation of Sp1 threonines 453 and 739, which are phosphorylated by ERK, decreased MBP transcriptional activity. Furthermore, we found that PKC regulates Sp1 phosphorylation only in differentiating OLs. In conclusion, our results indicate that, in OLs, Sp1 phosphorylation can be regulated by PKC-ERK pathways. This phosphorylation is important for MBP transcription and oligodendrocyte differentiation.
Guo
Lili
L
Division of Basic Biomedical Sciences, Sanford School of Medicine, University of South Dakota, Vermillion, South Dakota 57069, USA.
Eviatar-Ribak
Tamar
T
Miskimins
Robin
R
D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D047369:Chromatin Immunoprecipitation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D005786:Gene Expression Regulation; D004676:Myelin Basic Protein / Q000096:biosynthesis* / Q000235:genetics; D009836:Oligodendroglia / Q000166:cytology / Q000378:metabolism*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D016329:Sp1 Transcription Factor / Q000378:metabolism*; D014158:Transcription, Genetic / Q000235:genetics*
D004676:Myelin Basic Protein; D016329:Sp1 Transcription Factor; D011493:Protein Kinase C; D048049:Extracellular Signal-Regulated MAP Kinases