Interaction of glyceraldehyde-3-phosphate dehydrogenase with secondary and tertiary RNA structural elements of the hepatitis A virus 3′ translated and non-translated regions

Abstract

Proteins interacting with RNA structures at the 39 non-translated region (39NTR) of picornaviruses are probably important during viral RNA replication. We have shown previously that a dominant cellular cytoplasmic protein of 38 kDa (p38) interacts with the 39NTR and upstream regions of the hepatitis A virus (HAV) RNA (Kusov et al., J Virol 70, 1890-1897. Immunological and biochemical analyses of p38 have indicated that it is identical to GAPDH, which has previously been described as modulating translational regulation of the HAV RNA by interacting with the 59NTR (Schultz et al., J Biol Chem 271, 14134-14142, 1996). Three separate binding regions for GAPDH in the 39NTR and in the upstream 3D polymerase-coding region were identified. Structural analysis of these RNA regions by computer modelling and direct enzymatic cleavage suggested the presence of several AU-rich stem-loop structures having the potential for tertiary interactions. Binding of GAPDH to these structures was confirmed by RNA footprint analysis and resulted in the loss of double-stranded RNA regions. A different panel of RNA binding proteins (p28, p41 and p65) was detected in the ribosomal fractions of several cell lines (BSC-1, FRhK-4 and HeLa), whereas RNA binding of the GAPDH that was also present in these fractions was only marginal or absent.

Glyceraldehyde-3-Phosphate Dehydrogenase Regulates Endothelin-1 Expression by a Novel, Redox-Sensitive Mechanism Involving mRNA Stability

“…This effect was abolished when the protein was pretreated with oxidant GSSG, a condition shown to covalently modify Cys 152. Published works have already reported that binding of GAPDH to AU-rich stem-loop structures results in the loss of double-stranded-RNA regions (15,47). We hypothesize that GAPDH may function as an RNA anti-chaperone protein, promoting by RNA unwinding and hence facilitating the degradation of RNA by additional cellular components.…”

“…Previous reports of GAPDH binding to AU-rich RNAs have described rearrangements of the RNA structure, with the loss of double-stranded-RNA regions upon interaction (15,47). It is therefore plausible to hypothesize that GAPDH interaction with ET-1 mRNA facilitates the degradation of the mRNA by modification of the RNA structure at the 3Ј UTR, making it more susceptible to attack by cytosolic ribonucleases.…”

In Vitro and In Vivo Mutational Analysis of the 3′-Terminal Regions of Hepatitis E Virus Genomes and Replicons

“…The 3Ј NCRs of some viruses have been shown to interact with host proteins (4,28,40). Therefore, given the extent of differences between strains Sar55 and Meng in these regions, it appeared reasonable to determine whether the Sar/3Јswine chimera could infect pigs.…”

“…The 3Ј NCR of viruses has been reported to interact with host proteins (4,28,40), and since there was a 26% difference in nucleotide sequence in this region between strains Sar55 and Meng, it seemed possible that this region might be responsible for the ability of the swine virus to infect different species. To test this hypothesis, we made one virus pool containing HEV from a rhesus monkey that had been transfected with transcripts synthesized in vitro from pSar55 and another virus pool from a monkey that had been transfected with transcripts synthesized in vitro from pSar/3Јswine; we inoculated approximately 10 6 genomes of Sar55 or of Sar/3Јswine into swine.…”

Proteomic Characterization of Messenger Ribonucleoprotein Complexes Bound to Nontranslated or Translated Poly(A) mRNAs in the Rat Cerebral Cortex

“…The presence of two enzymes involved in glycolysis (phosphofructokinase C and glyceraldehyde-3-phosphate dehydrogenase) in an mRNP preparation was not expected but may support the assumption that there exists a link between glycolysis and the protein synthesis machinery (26). Thus the product of the phosphofructokinase, fructose 1,6-bisphosphate, acts as a stimulant of mRNA translation (27), whereas the glyceraldehyde-3-phosphate dehydrogenase may affect mRNA translation via its know ability to bind viral mRNAs (28,29).…”