Activation of the RegB endoribonuclease by the S1 ribosomal protein is due to cooperation between the S1 four C-terminal modules in a substrate-dependant manner.
The RegB protein, encoded by the T4 bacteriophage genome, is a ribonuclease involved in the inactivation of the phage early messenger RNAs. Its in vitro activity is very low but can be enhanced up to 100-fold in the presence of the ribosomal protein S1. The latter is made of six repeats of a conserved module found in many other proteins of RNA metabolism. Considering the difference between its size (556 amino acids) and that of several RegB substrates (10 nucleotides), we wondered whether all six modules are necessary for RegB activation. We studied the influence of twelve S1 fragments on the cleavage efficiency of three short substrates. RegB activation requires the cooperation of different sets of modules depending on the substrates. Two RNAs are quite well cleaved in the presence of the fragment formed by the fourth and fifth modules, whereas the third requires the presence of the four C-terminal domains. However, NMR interaction experiments showed that, despite these differences, the interactions of the substrates with either the bi- or tetra-modules are similar, suggesting a common interaction surface. In the case of the tetra-module the interactions involve all four domains, raising the question of the spatial organization of this region.
Bisaglia
Marco
M
ICSN-RMN, Ecole Polytechnique, 91128 Palaiseau, France and the Institut Jacques Monod, 1 Place Jussieu, 75005 Paris, France.
Laalami
Soumaya
S
Uzan
Marc
M
Bontems
Francois
F
D000595:Amino Acid Sequence; D004722:Endoribonucleases / Q000378:metabolism*; D004789:Enzyme Activation; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000494:pharmacology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000502:physiology*
D029968:Escherichia coli Proteins; D010446:Peptide Fragments; D012269:Ribosomal Proteins; C015494:ribosomal protein S1; D004722:Endoribonucleases; C407103:T4 RegB endoribonuclease