Colicin E5 is a ribonuclease that specifically cleaves tRNA(Tyr), tRNA(His), tRNA(Asn) and tRNA(Asp) of sensitive Escherichia coli cells by recognizing their anticodon sequences. Since all organisms possess universal anticodons of these tRNAs, colicin E5 was expected to potentially cleave eukaryotic tRNAs. Here, we expressed the active domain of colicin E5 (E5-CRD) in Saccharomyces cerevisiae and investigated its effects on growth. E5-CRD impaired growth of host cells by cleaving tRNA(Tyr), tRNA(His), tRNA(Asn) and tRNA(Asp) in S. cerevisiae, which is the same repertoire as that in E. coli. This activity of E5-CRD was inhibited by the co-expression of its cognate inhibitor (ImmE5). Notably, the growth impairment by E5-CRD was reversible; cells restored the colony-forming activity after suppression of the E5-CRD expression. This seems different from the sharp killing effect of E5-CRD on E. coli. These results may provide insights into the role and behaviour of cytosolic tRNAs on cell growth and proliferation.
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http://dx.doi.org/10.1093/jb/mvp004 | DOI Listing |
Protein Sci
January 2023
Department of Biology, College of Arts & Sciences, Saint Louis University, Saint Louis, Missouri, USA.
Detection of homologous relationships among proteins and understanding their mechanisms of diversification are major topics in the fields of protein science, bioinformatics, and phylogenetics. Recent developments in sequence/profile-based and structural similarity-based methods have greatly facilitated the unification and classification of many protein families into superfamilies or folds, yet many proteins remain unclassified in current protein databases. As one of the three earliest identified RNases in biology, ribonuclease T2, also known as RNase I in Escherichia coli, RNase Rh in fungi, or S-RNase in plant, is thought to be an ancient RNase family due to its widespread distribution and distinct structure.
View Article and Find Full Text PDFProc Natl Acad Sci U S A
September 2021
Robert F. Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, NY 14853;
As a common protein modification, asparagine-linked (linked) glycosylation has the capacity to greatly influence the biological and biophysical properties of proteins. However, the routine use of glycosylation as a strategy for engineering proteins with advantageous properties is limited by our inability to construct and screen large collections of glycoproteins for cataloguing the consequences of glycan installation. To address this challenge, we describe a combinatorial strategy termed shotgun scanning glycomutagenesis in which DNA libraries encoding all possible glycosylation site variants of a given protein are constructed and subsequently expressed in glycosylation-competent bacteria, thereby enabling rapid determination of glycosylatable sites in the protein.
View Article and Find Full Text PDFProteins
April 2021
Department of Biological Sciences, KAIST Institute for the Biocentury, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea.
We report the crystal structure of PYCH_01220, a hypothetical protein in Pyrococcus yayanosii CH1. This protein is composed of two domains, named Domain A and Domain B. While Domain B is not significantly homologous to known protein structures, Domain A is structurally analogous to the C-terminal ribonuclease domain of Escherichia coli colicin D.
View Article and Find Full Text PDFRNA Biol
August 2021
Department of Biotechnology, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan.
Colicin D is a plasmid-encoded bacteriocin that specifically cleaves tRNA of sensitive cells. has four isoaccepting tRNAs; the cleavage occurs at the 3' end of anticodon-loop, leading to translation impairment in the sensitive cells. tRNAs form a common L-shaped structure and have many conserved nucleotides that limit tRNA identity elements.
View Article and Find Full Text PDFStructure
November 2019
Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA. Electronic address:
In this issue of Structure, Gucinski et al. (2019) have described structural and enzymatic characterizations of two tRNase ribotoxins. The study significantly advances our understanding on the evolution and the mode of action of a group of ribotoxins that cleave the accepting stem of tRNAs for cell killing.
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