%0 Journal Article %1 wagner_archaeal_2007 %A Wagner, Steffen %A Klug, Gabriele %D 2007 %J The Journal of biological chemistry %K Amino_Acid_Sequence Archaeal Archaeal_Proteins Base_Sequence Cells Cultured Eukaryotic_Cells Genetic_Vectors Halobacterium IFZ Ligands Lysine Messenger Molecular_Sequence_Data Nucleic_Acid Nucleic_Acid_Conformation Peptide_Initiation_Factors Post-Translational Protein_Processing RNA RNA-Binding_Proteins Ribonucleases Sequence_Homology %P 13966-76 %T An archaeal protein with homology to the eukaryotic translation initiation factor 5A shows ribonucleolytic activity %V 282 %X To identify proteins that are involved in RNA degradation and processing in Halobacterium sp. NRC-1, we purified proteins with RNA-degrading activity by classical biochemical techniques. One of these proteins showed strong homology to the eukaryotic initiation factor 5A (eIF-5A) and was accordingly named archaeal initiation factor 5A (aIF-5A). Eukaryotic IF-5A is known to be involved in mRNA turnover and to bind RNA. Hypusination of eIF-5A is required for sequence-specific binding of RNA. This unique post-translational modification is restricted to Eukarya and Archaea. The exact function of eIF-5A in RNA turnover remained obscure. Here we show for the first time that aIF-5A from Halobacterium sp. NRC-1 exhibits RNA cleavage activity, preferentially cleaving adjacent to A nucleotides. Detectable RNA binding could be shown for aIF-5A purified from Halobacterium sp. NRC-1 but not from Escherichia coli, while both proteins possess RNA cleavage activity, indicating that hypusination of aIF-5A is required for RNA binding but not for its RNA cleavage activity. Furthermore, we show that the hypusinated form of eIF-5A also shows RNase activity while the unmodified protein does not. Charged amino acids in the N-terminal domain of aIF-5A as well as in the C-terminal domain, which is highly similar to the cold shock protein A (CspA), an RNA chaperone of E. coli, are important for RNA cleavage activity. Moreover our results reveal that activity of aIF-5A depends on its oligomeric state.

@article{wagner_archaeal_2007, abstract = {To identify proteins that are involved in RNA degradation and processing in Halobacterium sp. NRC-1, we purified proteins with RNA-degrading activity by classical biochemical techniques. One of these proteins showed strong homology to the eukaryotic initiation factor 5A (eIF-5A) and was accordingly named archaeal initiation factor 5A (aIF-5A). Eukaryotic IF-5A is known to be involved in mRNA turnover and to bind RNA. Hypusination of eIF-5A is required for sequence-specific binding of RNA. This unique post-translational modification is restricted to Eukarya and Archaea. The exact function of eIF-5A in RNA turnover remained obscure. Here we show for the first time that aIF-5A from Halobacterium sp. NRC-1 exhibits RNA cleavage activity, preferentially cleaving adjacent to A nucleotides. Detectable RNA binding could be shown for aIF-5A purified from Halobacterium sp. NRC-1 but not from Escherichia coli, while both proteins possess RNA cleavage activity, indicating that hypusination of aIF-5A is required for RNA binding but not for its RNA cleavage activity. Furthermore, we show that the hypusinated form of eIF-5A also shows RNase activity while the unmodified protein does not. Charged amino acids in the N-terminal domain of aIF-5A as well as in the C-terminal domain, which is highly similar to the cold shock protein A (CspA), an RNA chaperone of E. coli, are important for RNA cleavage activity. Moreover our results reveal that activity of aIF-5A depends on its oligomeric state.}, added-at = {2008-05-07T15:52:44.000+0200}, author = {Wagner, Steffen and Klug, Gabriele}, biburl = {https://www.bibsonomy.org/bibtex/2dca52bc2607846f74bf0d7984035d738/mikromolbio}, interhash = {e3c0b8dae3587b5cbe7dbe1400082dd4}, intrahash = {dca52bc2607846f74bf0d7984035d738}, issn = {00219258}, journal = {The Journal of biological chemistry}, keywords = {Amino_Acid_Sequence Archaeal Archaeal_Proteins Base_Sequence Cells Cultured Eukaryotic_Cells Genetic_Vectors Halobacterium IFZ Ligands Lysine Messenger Molecular_Sequence_Data Nucleic_Acid Nucleic_Acid_Conformation Peptide_Initiation_Factors Post-Translational Protein_Processing RNA RNA-Binding_Proteins Ribonucleases Sequence_Homology}, month = May, note = {PMID: 17369252}, pages = {13966-76}, timestamp = {2008-07-18T14:09:56.000+0200}, title = {An archaeal protein with homology to the eukaryotic translation initiation factor 5A shows ribonucleolytic activity}, volume = 282, year = 2007 }