Details

Autor(en) / Beteiligte

• Nagy, Eszter
• Henics, Tamás
• Eckert, Mária
• Miseta, Attila
• Lightowlers, Robert N.
• Kellermayer, Miklós

Titel

Identification of the NAD+-Binding Fold of Glyceraldehyde-3-Phosphate Dehydrogenase as a Novel RNA-Binding Domain

Ist Teil von

• Biochemical and biophysical research communications, 2000-08, Vol.275 (2), p.253-260

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2000

Quelle

Elsevier Journal Backfiles on ScienceDirect (DFG Nationallizenzen)

Beschreibungen/Notizen

• There is growing evidence that metabolic enzymes may act as multifunctional proteins performing diverse roles in cellular metabolism. Among these functions are the RNA-binding activities of NAD+-dependent dehydrogenases. Previously, we have characterized the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an RNA-binding protein with preference to adenine-uracil-rich sequences. In this study, we used GST-GAPDH fusion proteins generated by deletion mutagenesis to search for the RNA binding domain. We established that the N-terminal 43 amino acid residues of GAPDH, which correspond to the first mononucleotide-binding domain of the NAD+-binding fold is sufficient to confer RNA-binding. We also provide evidence that this single domain, although it retains most of the RNA-binding activity, loses sequence specificity. Our results suggest a molecular basis for RNA-recognition by NAD+-dependent dehydrogenases and (di)nucleotide-binding metabolic enzymes that had been reported to have RNA-binding activity with different specificity. To support this prediction we also identified other members of the family of NAD+-dependent dehydrogenases with no previous history of nucleic acid binding as RNA binding proteins in vitro. Based on our findings we propose the addition of the NAD+-binding domain to the list of RNA binding domains/motifs.