Our studies demonstrate that the binding of P-site tRNA to empty or programmed ribosome leads to inhibition of ribosome’s chaperoning ability. Studies using antibiotics that act as PTC substrate analogs indicated that the interaction between 39-CCA end of Psite tRNA with domain V of 23S rRNA of the ribosome is responsible for the observed inhibition. Ribosomal ligands, targeted to bind to specific sites on the PTC were used in this study. A comparison of the relative doses of these ligands to titrate out ribosome’s chaperoning ability, in presence of appropriate internal controls, form the basis of the conclusions presented here. This study relies on the optimum conditions for binding of tRNAs or antibiotics to the ribosome as reported in literature. Further studies are necessary to determine the exact occupancy of the ribosome with its substrates under our experimental conditions and to correlate the inhibition of ribosome’s chaperoning function with the concentration of the ribosome substrate complex. Mutagenesis studies presented here have identified two nucleotides that are necessary for tRNA binding, U2585 and G2252 to be important for chaperoning ability of bdV RNA. Both these mutants showed a deficiency in the release of the bound protein, which is an important step in the mechanism of bDV RNA mediated protein folding. Thus, whether the presence of a P-site tRNA prevents access of the refolding protein to the domain V of 23S rRNA of refolding process also needs to be further ascertained. The discussion of some recent observations are relevant in context of the above studies that imply that actively translating ribosomes with a tRNA positioned at the P-site would be unable to perform their chaperoning function. Firstly, the question as to whether the ribosome acts as a chaperone by trans or cis mechanism still remains to be answered. In the cismechanism, the conflict of the two concepts, the ribosome exit tunnel and protein folding on the PTC of the 50S subunit arises where indeed the question of how the nascent protein associates with the PTC after being released from the exit site of the tunnel needs to be further resolved. In this perspective, whether the chaperoning ability of the translating ribosome is activated after release factor mediated termination of polypeptide synthesis i.e. when the deacylated P-site tRNA moves to the P/E state, requires further investigation. Secondly, recent in vivo studies using PSI+ and Ure 3 strains of yeast Saccharomyces cerevisiae have identified the ribosomal RNA as target for two antiprion drugs, 6AP and GA. It was also demonstrated that these compounds selectively inhibit the protein folding ability of the ribosome and that the competitive obstruction of the protein binding sites of 23S rRNA by 6AP forms the basis of the inhibitory effect of the drug. These studies imply that the protein folding ability of the ribosome might also have impact on MK-4827 diverse cellular activities. Thirdly, our studies also raise the possibility that the ribosomes not engaged in active translation or its isolated 50S subunit present in the cell would be involved in assisting cellular protein folding.