Archaebacterial Genomes: Eubacterial Form and Eukaryotic Content

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Archaebacterial Genomes: Eubacterial Form and Eukaryotic Content Archaebacterial genomes: eubacterial form and eukaryotic content Patrick J Keeling, Robert LCharlebois and W Ford Doolittle Dalhousie University, Halifax and University of Ottawa, Ottawa, Canada Since the recognition of the uniqueness and coherence of the archaebacteria (sometimes called Archaea), our perception of their role in early evolution has been modified repeatedly. The deluge of sequence data and rapidly improving molecular systematic methods have combined with a better understanding of archaebacterial molecular biology to describe a group that in some ways appears to be very similar to the eubacteria, though in others is more like the eukaryotes. The structure and contents of archaebacterial genomes are examined here, with an eye to their meaning in terms of the evolution of cell structure and function. Current Opinion in Genetics and Development 1994, 4:816-822 Introduction this review we will focus on more recent discoveries and try to assimilate these observations with phylogeny. As molecular phylogenetics advances, it provides evolu- tionary biologists with a f&rework on which to assem- ble a coherent picture of the history of life. Two un- Archaebacterial genome design expected discoveries that emerged from sequence com- parisons have had particularly far-reaching effects on our At present, seven archaebacterial genomes have been view of early evolution; both concern the archaebacteria. physically mapped. In each case, the genome is com- The first, on the basis of ribosomal RNA (rRNA) se- posed of a single circular chromosome that falls within quence information, and now extensively supported by a the size range observed in the eubacteria [5-11,12”,13*] variety of other data, was the finding that the prokaryotes (Table 1). In several instances, extrachromosomal ele- are actually composed of two distinct groups, eubacteria ments have also been identified [14]; in some halophiles, and archaebacteria, as distant horn one another as either plasmid and megaplasmid (up to 700 kb) DNA makes are f%om the eukaryotes [l]. The second, equally excit- up a substantial portion of the genome [15]. Halophile ing discovery is that the eubacteria branched first from plasmids and chromosomes bear scores of transposable the universal tree, so that the archaebacteria and eukary- elements, which give rise to high frequencies of ge- otes seem to be sister groups [2]. The significance of this netic instability as revealed by spontaneous mutations and second conclusion (which, although increasingly popu- local genetic rearrangements detectable through South- lar, does need confirmation from additional data sets) is ern analysis [16]. This phenomenon can be observed on that it allows us to propose specific arguments about the the time scale of laboratory experiments, but compara- nature of both the prokaryotic ancestor of the eukary- tive mapping indicates that in evolutionary time, chro- otes and the last common ancestor of all extant life on mosomal order is remarkably conserved in halobacteria the basis of characters shared between pairs of taxa. (P Lopez-Garcia, A St Jean, R Amils, R Charlebois, un- published data; N Hackett, personal communication). As our understanding of the archaebacteria on the Forces must exist that can effectively oppose the pow- molecular level develops, it is becoming clear that they erful destabilizing pressures present in the halobacterial share a number of features in overall genome design and genome. gene organization with the eubacteria. These traits can then be said most parsimoniously to be ancestral to all Archaebacterial and eubacterial genomes are similar in life. Many other characters, particularly those involved size and structure, both being small, compact, and cir- with gene expression, are shared between the archae- cular. If the rooting of the universal tree by Iwabe [2] is bacteria and eukaryotes; these can be either primitive correct, it is reasonable to suppose that this design is an- or derived, but in both cases help us to understand the cestral to all extant life, although we know too few of the order of evolutionary events at or near the origin of the details of genome construction to be certain that the trait eukaryotes. Detailed reviews of archaebacterial molecu- could not have evolved twice independently. To describe lar biology are available in two recent books [3,4], so in the nature of the primitive genome more accurately, we Abbreviation snoRNA-small nucleolar RNA. 816 0 Current Biology Ltd ISSN 0959-437X Archaebacterial genomes Keeling, Charlebois and Doolittle 817 order S12-S7-EF-G-EF-Tu for both M. uunnielii and Table 1. Archaebacterial genome sizes as determined by mapping. E. coli [28]). In one case, the M. vannielii Ll operon, the similarity can be extended to the regulation of ex- Species Cenome size Method References pression, as it has been shown that the Ll protein in M. vannielii represses translation of its own mRNA by bind- Mefhanococcus voltae 1900 PFC I71 ing to a site that resembles its binding site on the 23s tialobacrerium halobium 2400 PFC 1101 rRNA [25**]. The same autoregulation is seen in E. coli, Halobacterium sp. CR6 2472 Cloning [12-l differing only in the position of the binding site on the Haloferax volcanii 4140 Cloning 161 mRNA. Haloferax mediterranei 3840 PFG l8,13*1 Thermococcus celer 1890 PFG 151 Enough data may not be available to say that gene or- Sulfolobus acidocaldarius 2760 PFG I91 der is conserved over even longer distances in eubacterial and archaebacterial chromosomes, but enough is known Summary of archaebacterial genomes that have been mapped. Each to begin to assemble some intriguing correlations. The of these gendmes is within the range of genome size documented in spectinomycin operon is immediately downstream of the eubacteria and all finished maps are circular. PFG, pulsed-field gel the SlO operon in both E. coli and M. vannielii [17], electrophoresis. and the streptomycin operon is followed by the SlO operon in Themotogu maritima and by the SlO protein (the first reading tie in the SlO operon) in three d& need more comparative data and a better understanding ferent archaebacteria [28], as well as in the cyanobac- of archaebacterial genomes Corn a functional standpoint; terium Spinrlina platensis [29]. The current best guess for instance, the mechanisms of replication initiation, is that in the ancestor of archaebacteria wd eubac- termination, and chromosomal segregation need to be teria, these three operons followed one another in the examined. order streptomycin-SlO-spectinomycin, with the genes in each being similar to those found commonly in con- Archaebacterial gene organization temporary operons. In general, the structure of archaebacterial genes closely The transition to eukaryotes resembles that of eubacteria [15,17]. They are organized into co-transcribed units, or operons, that are expressed A particularly close afinity between archaebacteria and as long non-capped mRNAs with only short bacterial- eukaryotes has been long suspected, but for the most part like poly(A) tails [18]. Protein-coding genes of the ar- on vague grounds - resistance to certain drugs, lack chaebacteria are not disrupted by introns of any type, of peptidoglycan, or the occurrence of tRNA introns other than some spliced at the protein level [19,20], and [30]. Recent work, however, has elucidated a ham&l to date no intron resembling the nuclear spliceosomal of molecular similarities that argue unambiguously for a type has been identified. Introns have been found in shared inheritance with the eukaryotes, three of which 16S, 23s. and tRNA genes; these are of a novel type, are reviewed here. dependent upon a defined secondary structure at the intron/exon boundary and an unidentified enzymatic One of the strongest similarities is in transcription mech- activity [21]. Some of these introns have been found to anisms, now known to dif&r fundamentally &om the contain open reading frames related to homing endonu- homologous eubacterial process. Evidence for this simi- cleases first characterized in group I self-splicing introns larity has been accumulating for a number of years, be- of mitochondria and eubacteriophages [22*]. ginning with the observation that archaebacterial RNA polymerases are multisubunit complexes, unlike the sim- The degree of similarity between archaebacterial and ple three or four subunit eubacterial enzyme [31,32], eubacterial operons or operon clusters is best exemplified and that the consensus archaebacterial promoter bears by a number of archaebacterial operons that contain the a closer resemblance to the eukaryotic TATA box than same genes in the same order as their eubacterial homo- to eubacterial promoter sequences [33]. The depth of logs (reviewed in [23]; see Fig. 1). These include the this similarity was further revealed when the existence of RNA polymerase operon and ribosomal protein gene an archaebacterial homolog of the eukaryotic transcrip- clusters, which are homologous to the Escherichiu colt’ tion &ctor TFIIB was discovered by a database search Lll and LlO clusters (four of four genes in the same [34,35*], and by wo reports of a gene that encodes order in two archaebacteria and E. coli [24,25”]). the a protein closely resembling both TFIIS and a subunit spectinomycin operon (for which Methunococcusvunnielii of eukaryotic RNA polymerases I and II [36’,37*]. and Sulfolobus ucidoculduriusshow eleven and nine, respec- Moreover, work by two independent investigators has tively, of the eleven E. coli genes in the same order, now shown that archaebacterial genomes also encode although the archaebacteria each have three ‘extras’, TFIID, or the TATA-binding protein [38”,39”], and which are homologs of eukaryotic ribosomal protein that TFIID recognizes archaebacterial promoters and genes [24,26]), the SlO operon (seven genes in a stretch directs the binding of TFIIB [39”]. Unlike their eubac- of seven in the same order in halobacteria and E. coli terial counterparts, eukaryotic RNA polymerases alone [27]), and the streptomycin operon (four genes in the cannot recognize promoters; instead, the assembly of the 818 Genomes and evolution ‘Lll , LlO Clusters E.
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