Thermodynamics of the fragile X mental retardation protein RGG box interactions with G quartet forming RNA.
Fragile X syndrome, the most common form of inherited mental retardation, is the result of an unstable expansion of a CGG trinucleotide repeat in the 5' UTR of the fragile X mental retardation-1 (FMR1) gene. The abnormal hypermethylation of the expanded CGG repeats causes the transcriptional silencing of the FMR1 gene and, consequently, the loss of the fragile X mental retardation protein (FMRP). FMRP is an RNA binding protein that binds to G quartet forming RNA using its RGG box motif. In this study we have performed a thermodynamic analysis of the interactions between the FMRP RGG box domain and Sc1, an RNA molecule which had been previously shown to be bound with high affinity by both the full-length FMRP and by its RGG box domain. We have determined that the association between the FMRP RGG box and Sc1 RNA is dominated by hydrophobic and hydrogen bond interactions, with minor contributions from electrostatic interactions, and that the FMRP RGG box binding increases the stability of the G quartet RNA structure significantly. Interestingly, we found that the G quartet recognition is necessary but not sufficient for the FMRP RGG box binding to this RNA target, indicating that additional interactions of the peptide, possibly with the stem and/or stem-G quartet junction region, are required. Our results also indicate that the G quartet RNA recognition is not a general feature of the RGG box motif but rather carries some sequence, protein and/or RNA, specificity.
Zanotti
Kimberly J
KJ
Department of Chemistry and Biochemistry, Duquesne University, Pittsburgh, Pennsylvania 15282, USA.
Lackey
Patrick E
PE
Evans
Genevieve L
GL
Mihailescu
Mihaela-Rita
MR
D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000737:chemistry*; D005998:Glycine / Q000737:chemistry; D006801:Humans; D009690:Nucleic Acid Conformation; D017434:Protein Structure, Tertiary / Q000235:genetics; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
D001120:Arginine; D051860:Fragile X Messenger Ribonucleoprotein 1; D005998:Glycine; D012313:RNA; D016601:RNA-Binding Proteins