Circularization restores signal recognition particle RNA functionality in Thermoproteus

Signal recognition particles (SRPs) are universal ribonucleoprotein complexes found in all three domains of life that direct the cellular traffic and secretion of proteins. These complexes consist of SRP proteins and a single, highly structured SRP RNA. Canonical SRP RNA genes have not been identified for some Thermoproteus species even though they contain SRP19 and SRP54 proteins. Here, we show that genome rearrangement events in Thermoproteus tenax created a permuted SRP RNA gene. The 5'- and 3'-termini of this SRP RNA are located close to a functionally important loop present in all known SRP RNAs. RNA-Seq analyses revealed that these termini are ligated together to generate circular SRP RNA molecules that can bind to SRP19 and SRP54. The circularization site is processed by the tRNA splicing endonuclease. This moonlighting activity of the tRNA splicing machinery permits the permutation of the SRP RNA and creates highly stable and functional circular RNA molecules.

Thermoproteus thermophilus sp. nov., a hyperthermophilic crenarchaeon isolated from solfataric soil

A hyperthermophilic crenarchaeon, designated strain CBA1502T, was isolated from volcanic soil in the Mayon volcano in the Philippines. The 16S rRNA gene sequence of strain CBA1502T was most closely related to that of Thermoproteus uzoniensis DSM 5263T (99.2 % similarity) and Thermoproteus tenax Kra 1T (99.0 %). The organism grew at 75–90 °C and pH 4.0–6.0 and in the presence of 0–0.5 % (w/v) NaCl, with optimal growth at 85 °C and pH 5.0. Strain CBA1502T utilized d-arabinose, beef extract, Casamino acids, formate, fumarate, peptone, pyruvate, trimethylamine and yeast extract as energy substrates, and d-arabinose, formate, pyruvate and yeast extract as carbon sources. Fumarate, sulfate, sulfur and thiosulfate functioned as electron acceptors, but not ferric chloride, nitrate, malate or oxidized glutathione. DNA–DNA hybridization studies showed that there was less than 46.1 % relatedness between strain CBA1502T and other members of the genus Thermoproteus. The DNA G+C content of strain CBA1502T was 62.0 mol%. We conclude that, according to its phylogenetic, phenotypic and genotypic characteristics, strain CBA1502T represents a novel species of the genus Thermoproteus, and propose the name Thermoproteus thermophilus sp. nov., with the type strain CBA1502T ( = ATCC BAA-2416T = JCM 17229T).

Extensive Lysine Methylation in Hyperthermophilic Crenarchaea: Potential Implications for Protein Stability and Recombinant Enzymes

In eukarya and bacteria, lysine methylation is relatively rare and is catalysed by sequence-specific lysine methyltransferases that typically have only a single-protein target. Using RNA polymerase purified from the thermophilic crenarchaeumSulfolobus solfataricus, we identified 21 methyllysines distributed across 9 subunits of the enzyme. The modified lysines were predominantly inα-helices and showed no conserved sequence context. A limited survey of theThermoproteus tenaxproteome revealed widespread modification with 52 methyllysines in 30 different proteins. These observations suggest the presence of an unusual lysine methyltransferase with relaxed specificity in the crenarchaea. Since lysine methylation is known to enhance protein thermostability, this may be an adaptation to a thermophilic lifestyle. The implications of this modification for studies and applications of recombinant crenarchaeal enzymes are discussed.

Genome Sequence of Thermofilum pendens Reveals an Exceptional Loss of Biosynthetic Pathways without Genome Reduction

ABSTRACT We report the complete genome of Thermofilum pendens, a deeply branching, hyperthermophilic member of the order Thermoproteales in the archaeal kingdom Crenarchaeota. T. pendens is a sulfur-dependent, anaerobic heterotroph isolated from a solfatara in Iceland. It is an extracellular commensal, requiring an extract of Thermoproteus tenax for growth, and the genome sequence reveals that biosynthetic pathways for purines, most amino acids, and most cofactors are absent. In fact, T. pendens has fewer biosynthetic enzymes than obligate intracellular parasites, although it does not display other features that are common among obligate parasites and thus does not appear to be in the process of becoming a parasite. It appears that T. pendens has adapted to life in an environment rich in nutrients. T. pendens was known previously to utilize peptides as an energy source, but the genome revealed a substantial ability to grow on carbohydrates. T. pendens is the first crenarchaeote and only the second archaeon found to have a transporter of the phosphotransferase system. In addition to fermentation, T. pendens may obtain energy from sulfur reduction with hydrogen and formate as electron donors. It may also be capable of sulfur-independent growth on formate with formate hydrogen lyase. Additional novel features are the presence of a monomethylamine:corrinoid methyltransferase, the first time that this enzyme has been found outside the Methanosarcinales, and the presence of a presenilin-related protein. The predicted highly expressed proteins do not include proteins encoded by housekeeping genes and instead include ABC transporters for carbohydrates and peptides and clustered regularly interspaced short palindromic repeat-associated proteins.

DNA Microarray Analysis of Central Carbohydrate Metabolism: Glycolytic/Gluconeogenic Carbon Switch in the Hyperthermophilic Crenarchaeum Thermoproteus tenax

ABSTRACT In order to unravel the role of regulation on transcript level in central carbohydrate metabolism (CCM) of Thermoproteus tenax, a focused DNA microarray was constructed by using 85 open reading frames involved in CCM. A transcriptional analysis comparing heterotrophic growth on glucose versus autotrophic growth on CO2-H2 was performed.