Abstract
The Escherichia coli rnt gene, which encodes the RNA-processing enzyme RNase T, is cotranscribed with a downstream gene. Complete sequencing of this gene indicates that its coding region encompasses 1,538 amino acids, making it the longest known protein in E. coli. The gene (tentatively termed lhr for long helicase related) contains the seven conserved motifs of the DNA and RNA helicase superfamily II. An approximately 170-kDa protein is observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 35S-labeled extracts prepared from cells in which lhr is under the control of an induced T7 promoter. This protein is absent when lhr is interrupted or when no plasmid is present. Downstream of lhr is the C-terminal region of a convergent gene with homology to glutaredoxin. Interruptions of chromosomal lhr at two different positions within the gene do not affect the growth of E. coli at various temperatures in rich or minimal medium, indicating that lhr is not essential for usual laboratory growth. lhr interruption also has no effect on anaerobic growth. In addition, cells lacking Lhr recover normally from starvation, plate phage normally, and display normal sensitivities to UV irradiation and H2O2. Southern analysis showed that no other gene closely related to lhr is present on the E. coli chromosome. These data expand the known size range of E. coli proteins and suggest that very large helicases are present in this organism.
MeSH Terms
Amino Acid Sequence
Bacterial Proteins/genetics
Base Sequence
Conserved Sequence/genetics
DNA Helicases/biosynthesis,chemistry,genetics
Escherichia coli/enzymology,genetics,growth & development
Escherichia coli Proteins
Exoribonucleases/genetics
Genes, Bacterial/genetics
Molecular Sequence Data
Molecular Weight
Mutation
Restriction Mapping
Sequence Alignment
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Chemicals
Bacterial Proteins
Escherichia coli Proteins
Exoribonucleases
exoribonuclease T
DNA Helicases
Lhr protein, E coli
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Reuven N B
Department of Biochemistry, University of Connecticut Health Center, Farmington 06032, USA.
Koonin E V
Rudd K E
Deutscher M P
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