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W Zillig

Publications and source records attributed to W Zillig.

At least 55 records · Page 3Linked to original sources

Nucleotide sequence of the genes encoding proline tRNA(UGG) and threonine tRNA(GGU) and consensus promoter model of Thermococcus celer.

The nucleotide sequences of the genes encoding tRNA(Pro)(UGG) and tRNA(Thr)(GGU) from the extremely thermophilic archaeon (archaebacterium) Thermococcus celer have been determined. A consensus promoter model was deduced from the comparison of the upstream regions of several stable RNA genes with S1-mapped promoter regions of genes coding for ribosomal proteins and DNA-dependent RNA polymerase components.

Archaea↗

In vivo and in vitro analysis of transcription of the L region from the Halobacterium salinarium phage phi H: definition of a repressor-enhancing gene.

The 12-kb L region of the Halobacterium salinarium phage phi H is able to replicate as a plasmid, conferring a certain immunity to the host cell. We show here that the whole region is utilised for transcription at one stage or another in the phage life cycle. The DNA segment between the lytic transcripts T4 and T1 is shown to be constitutively transcribed. The effects of transcription on immunity were investigated using transformation experiments with H. salinarium. In this way, we show in vivo that the immune transcript T9, which on its own has no influence on phage growth or immunity, has a co-operative effect on the phi H repressor. The immunity effects analysed are sufficient to account for the degree of immunity conferred by the L region.

Amino Acid Sequence↗

SSV1-encoded site-specific recombination system in Sulfolobus shibatae.

We present evidence for the existence of a conservative site-specific recombination system in Archaea by demonstrating integrative recombination of Sulfolobus shibatae virus SSV1 DNA with the host chromosome, catalysed by the SSV1-encoded integrase in vitro. The putative int gene of SSV1 was expressed in Escherichia coli yielding a protein of about 39 kDa. This protein alone efficiently recombined linear DNA substrates containing chromosomal (attA) and viral (attP) attachment sites; recombination with either negatively or positively supercoiled SSV1 DNA was less efficient. Intermolecular attA x attA and attP x attP recombination was also promoted by the SSV integrase. The invariant 44 bp "common attachment core" present in all att sites contained sufficient information to allow recombination, whilst the flanking sequences effected the efficiency. These features clearly distinguish the SSV1--encoded site--specific recombination system from others and make it suitable for the study of regulatory mechanisms of SSV1 genome--host chromosome interaction and investigations of the evolution of the recombination machinery.

Bacteriophages↗

Antisense RNA mediates transcriptional processing in an archaebacterium, indicating a novel kind of RNase activity.

Strains of the extremely halophilic archaebacterium Halobacterium salinarium that are lysogenic for the phage phi H produce an antisense RNA transcript complementary to the first 151 nucleotides (nt) of the early lytic phage transcript T1. This is the first case of antisense control of gene expression in an archaebacterium. We show through transformation of H. salinarium that the antisense RNA functions in trans, rendering the early lytic phage transcript T1 susceptible to specific cleavage by an unidentified RNase of unique endonucleolytic activity. The single-stranded ends of RNA are cut off at the ends of the 151 nt RNA duplex, removing the ribosomal binding sites from the first open reading frame of transcript T1 but without concomitant digestion of the products.

Amino Acid Sequence↗

Elements of an archaeal promoter defined by mutational analysis.

The sequence requirements for specific and efficient transcription from the 16S/23S rRNA promoter of Sulfolobus shibatae were analysed by point mutations and by cassette mutations using an in vitro transcription system. The examination of the box A-containing distal promoter element (DPE) showed the great importance of the TA sequence in the center of box A for transcription efficiency and the influence of the sequence upstream of box A on determining the distance between the DPE and the start site. In most positions of box A, replacement of the wild type bases by adenines or thymines are less detrimental than replacements by cytosines or guanines. The effectiveness of the proximal promoter element (PPE) was not merely determined by its high A + T content but appeared to be directly related to its nucleotide sequence. At the start site a pyrimidine/purine (py/pu) sequence was necessary for unambiguous initiation as shown by analysis of mutants where the wild type start base was replaced. The sequence of box A optimal for promoter function in vitro is identical to the consensus of 84 mapped archaeal promoter sequences.

Base Composition↗

The particle SSV1 from the extremely thermophilic archaeon Sulfolobus is a virus: demonstration of infectivity and of transfection with viral DNA.

The lemon-shaped "virus-like" particle SSV1 produced by the thermophilic archaeon Sulfolobus shibatae has not previously been observed to infect any host. Using a plaque assay suitable for the extreme growth conditions of this archaeon, we have shown infection of Sulfolobus solfataricus by SSV1. Upon infection, the viral genome was always found integrated into a tRNA gene of the host chromosome, a situation similar to that in S. shibatae, proving that site-specific integration is involved in establishing the lysogenic state. As in S. shibatae, UV-irradiation of lysogenized S. solfataricus led to virus production apparently not accompanied by cell lysis. We have also demonstrated the efficient uptake of exogenous DNA and its expression in Sulfolobus by transfecting S. solfataricus with SSV1 DNA by electroporation. Transfection efficiencies of up to 10(6) transfectants per microgram of DNA were obtained.

Bacteriophages↗

Component H of the DNA-dependent RNA polymerases of Archaea is homologous to a subunit shared by the three eucaryal nuclear RNA polymerases.

The gene encoding component H of the DNA-dependent RNA polymerase (RNAP, EC 2.7.7.6) of Sulfolobus acidocaldarius has been identified by comparison of the amino acid sequence with the derived amino acid sequence of an open reading frame (ORF88) in the RNAP operon. Corresponding genes were identified in Halobacterium halobium and were cloned and sequenced from Thermococcus celer and Methanococcus vannielii. All these rpoH genes are situated between the promoters of the RNAP operons and the corresponding rpoB and rpoB2 genes. The archaeal H subunits show high sequence similarity to each other and to the C-terminal portions of the largest of four subunits shared by all three specialized nuclear RNAPs. These correlations are further evidence for the striking similarity between archaeal and eucaryal RNAP structures and transcription systems.

Amino Acid Sequence↗

In vivo studies on the effects of immunity genes on early lytic transcription in the Halobacterium salinarium phage phi H.

We have studied in vivo the effects of putative immunity genes on the expression of an early lytic gene of the Halobacterium salinarium phage. phi H. We transformed an H. salinarium host with DNA coding for a putative repressor gene, the transcript of which has been designated T6. We show that, in vivo, this gene specifically shuts off production of the early lytic transcript T4. A construct carrying the DNA transcribed as T4, but without its putative repressor binding sequences, shows T4 transcription enhanced to a level comparable to that observed in lytic growth of mutant phages capable of growing on immune H. salinarium strains. This transcript is insensitive to the action of the T6 product. The product of this 'unrepressed' T4 transcript is able to complement in trans the repressed T4 on superinfecting phi H-sensitive phages, allowing these to grow on a strain containing the repressor gene. It has, however, no effect on the production of repressor. We also mapped the start and end points of two other transcripts, T9 and T10, which are expressed only in the lysogenic state by cells immune to superinfection by phage, cloned the coding DNA and used it to transform H. salinarium. This DNA, though transcribed by the transformants, has no detectable effect on the cells, which remain susceptible to phage infection.

Bacteriophages↗

The immunity-conferring plasmid p phi HL from the Halobacterium salinarium phage phi H: nucleotide sequence and transcription.

The complete nucleotide sequence of the plasmid p phi HL, composing the central 12,041-bp L-region from the temperate phage phi H of Halobacterium salinarium is presented. Transcripts mapped to the p phi HL and the L-region produced under immune conditions, under lytic growth or constitutively, are described. The sequences upstream of the transcription start points show homology to the consensus sequence for archaeal (formerly archaebacterial) promoters. Lytic transcription is shown to be strictly time-dependent, with an early gene product required for the expression of late genes.

Bacteriophages↗

Heterologous in vitro transcription from two archaebacterial promoters.

A cell-free extract of Sulfolobus shibatae is able to specifically initiate transcription in vitro at the promoter of the plasmid-encoded gene for the major gas vesicle protein of Halobacterium halobium and at the promoter for the transcript T4 of the temperate H. halobium phage phi H. The corresponding promoter from the virulent phage mutant phi HL1 yields enhanced transcription in the heterologous system, in agreement with strongly increased in vivo expression.

Archaea↗

Comparative biochemistry of Archaea and Bacteria.

This review compares exemplary molecular and metabolic features of Archaea and Bacteria in terms of phylogenetic aspects. The results of the comparison confirm the coherence of the Archaea as postulated by Woese. Archaea and Bacteria share many basic features of their genetic machinery and their central metabolism. Similarities and distinctions allow projections regarding the nature of the common ancestor and the process of lineage diversification.

Archaea↗