Ribonuclease PH interacts with an acidic ribonuclease E site through a basic 80-amino acid domain
Víctor Pérez Medina Martínez, Gianni Dehò, Robert W. Simons, Jaime Garcı́a-Mena
Abstract
Víctor Pérez Medina Martínez, Gianni Dehò, Robert W. Simons, Jaime Garcı́a-Mena
Abstract
In this work, we characterize the domains for the in vivo interaction between ribonuclease E (RNase E) and ribonuclease PH (RNase PH). We initially explored the interaction using pull-down assays with full wild-type proteins expressed from a chromosomal monocopy gene. Once the interaction was confirmed, we narrowed down the sites of interaction in each enzyme to an acidic 16-amino acid region in the carboxy-terminal domain of RNase E and a basic 80-amino acid region in RNase PH including an α3 helix. Our results suggest two novel functional domains of interaction between ribonucleases.
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In this work, we characterize the domains for the in vivo interaction between ribonuclease E (RNase E) and ribonuclease PH (RNase PH). We initially explored the interaction using pull-down assays with full wild-type proteins expressed from a chromosomal monocopy gene. Once the interaction was confirmed, we narrowed down the sites of interaction in each enzyme to an acidic 16-amino acid region in the carboxy-terminal domain of RNase E and a basic 80-amino acid region in RNase PH including an α3 helix. Our results suggest two novel functional domains of interaction between ribonucleases.
Key concepts: Ribonuclease, RNase P, S-tag, Ribonuclease III, RNase PH, Amino acid, RNase H, Biochemistry