A novel protein, Xenopus p20, influences the stability of MeCP2 through direct interaction.
MeCP2 is the founder member of a family of methyl-CpG-binding proteins able to repress transcription from methylated DNA. To date, MeCP2 action seems to involve the delivery on modified DNA of histone deacetylase activity, followed by histone methylating activity. It has been recently demonstrated that MECP2 mutations cause Rett syndrome, a childhood neurological disorder that represents one of the most common causes of mental retardation in females. Here we show that a novel Xenopus laevis protein of 20 kDa, p20, is able to interact in vivo and in vitro with MeCP2. The p20 sequence revealed that it belongs to the family of the WAP (whey acidic protein) proteins, often functioning as a protease inhibitor. Therefore, we asked whether the p20 can influence the MeCP2 half-life. We demonstrate that, indeed, the xp20 not only can significantly increase the stability of an exogenously expressed MeCP2 in Xenopus oocytes but also can stabilize the human endogenous MeCP2. The capability of the mammalian methyl-CpG-binding protein to interact with p20 is confirmed by co-immunoprecipitation experiments performed overexpressing the WAP protein. Glutathione S-transferase pull-down assays reveal that the MeCP2 residues localized between the methyl-binding domain and the transcriptional repression domain is the primary interaction surface. Our data suggest that regulation of MeCP2 metabolism might be of relevant importance; in accordance with this, previous results have shown that some Rett syndrome mutations are characterized by a decrease in MeCP2 stability.
Carro
Stella
S
Dipartimento di Biologia Strutturale e Funzionale, Università degli Studi dell'Insubria, via Alberto da Giussano 12, 21052 Busto Arsizio, Italy.
Bergo
Anna
A
Mengoni
Mauro
M
Bachi
Angela
A
Badaracco
Gianfranco
G
Kilstrup-Nielsen
Charlotte
C
Landsberger
Nicoletta
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D051783:Methyl-CpG-Binding Protein 2; D008894:Milk Proteins / Q000494:pharmacology*; D008969:Molecular Sequence Data; D012097:Repressor Proteins; D015518:Rett Syndrome; D029867:Xenopus Proteins / Q000494:pharmacology*; D014982:Xenopus laevis
D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; C497395:MECP2 protein, Xenopus; C497392:MECP2 protein, human; D051783:Methyl-CpG-Binding Protein 2; D008894:Milk Proteins; D012097:Repressor Proteins; D029867:Xenopus Proteins; C024471:whey acidic proteins