Eukaryotic Features Research Articles

The complete nucleotide sequence of the tobacco chloroplast genome: its gene organization and expression

The complete nucleotide sequence (155 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA has been determined. It contains two copies of an identical 25 339 bp inverted repeat, which are separated by a 86 684 bp and a 18 482 bp single-copy region. The genes for 4 different rRNAs, 30 different tRNAs, 39 different proteins and 11 other predicted protein coding genes have been located. Among them, 15 genes contain introns. Blot hybridization revealed that all rRNA and tRNA genes and 27 protein genes so far analysed are transcribed in the chloroplast and that primary transcripts of the split genes hitherto examined are spliced. Five sequences coding for proteins homologous to components of the respiratory-chain NADH dehydrogenase from human mitochondria have been found. The 30 tRNAs predicted from their genes are sufficient to read all codons if the ;two out of three' and ;U:N wobble' mechanisms operate in the chloroplast. Two sequences which autonomously replicate in yeast have also been mapped. The sequence and expression analyses indicate both prokaryotic and eukaryotic features of the chloroplast genes.

Archaebacteria: the road to the universal ancestor.

AbstractLiving forms are now divided into three primary kingdoms, eubacteria, archaebacteria and eukaryotes. The archaebacteria demonstrate a blending of eubacterial and eukaryotic features and cast new perspectives on the nature of the universal ancestor.

Transformation of Schizosaccharomyces pombe by non-homologous, unstable integration of plasmids in the genome.

In the fission yeast, Schizosaccharomyces pombe, transformation with recombinant plasmids always results in a high proportion of mitotically unstable transformants. This suggested that specialised (ARS) sequences might not be required for autonomous replication of plasmids in S. pombe, contrary to the situation in Saccharomyces cerevisiae. We have shown that specialised ARS sequences, analogous to those in S. cerevisiae, do exist in S. pombe, supporting the view that ARS elements are a general feature of eukaryotes. In addition, there is a further mechanism of plasmid maintenance which involves homologous and non-homologous integration into, and excision from the genome.

Stimulation by silybin, a eukaryotic feature of archaebacterial RNA polymerases

Transcription of native DNA by DNA-dependent RNA polymerases of members of the thermoacidophilic branch of the archaebacteria like that of eukaryotic RNA polymerase A of rat liver, is stimulated by the flavonolignane derivative silybin. Eubacterial RNA polymerases are not affected. For both RNA polymerase A of rat liver and RNA polymerase of Thermoplasma acidophilum the stimulatory effect is confined to the enhancement of the elongation reaction suggesting a similar mechanism for both enzymes. These results further support a specific phylogenetic relationships between the archaebacteria and the eukaryotic cytoplasm.

Overlapping divergent genes in the maize chloroplast chromosome and in vitro transcription of the gene for tRNA His

In the presence of the S polypeptide, maize chloroplast DNA-dependent RNA polymerase preferentially transcribes sequences within the 2200-nucleotide-pair-long maize chloroplast chromosome fragment Eco [unk] from a supercoiled chimeric plasmid cloned in Escherichia coli [Jolly, S. O. & Bogorad, L. (1980) Proc. Natl. Acad. Sci. USA 77, 822-826]. Eco [unk] contains one gene for tRNA(His) and one for a 1.6-kilobase RNA that includes an open reading frame. These two genes overlap by at least a few nucleotides and are transcribed divergently from complementary DNA strands. This indicates possible transcriptional regulation of chloroplast DNA at the nucleotide level. The 5' end of tRNA(His) (G-U-G) isolated from maize chloroplasts is indistinguishable from that of the transcript produced from Eco [unk] in vitro by maize chloroplast DNA-dependent RNA polymerase. This purified system initiates RNA synthesis faithfully and exhibits preference for some chloroplast genes. Maize chloroplast DNA for tRNA(His) lacks the sequence C-C-A at its 3' terminus; it is presumably added post-transcriptionally. Maize tRNA(His) has both prokaryotic and eukaryotic features.

Nucleotide sequence of a spinach chloroplast threonine tRNA.

The nucleotide sequence of a spinach chloroplast threonine tRNA has been determined. This chloroplast threonine tRNA has been determined. This chloroplast threonine tRNA has 75 nucleotides, which is the same chain length as the recently determined threonine tRNA from yeast mitochondria. This contrasts with the 6 non-organelle threonine tRNAs sequenced to date, which are 76 nucleotides in length. However, other than this similarity in size, the chloropast tRNAThr has little similarlity to the yeast mitochondrial threonine tRNA, and shows essentially equal homology to both prokaryotic and eukaryotic feature of sequence homology to other threonine tRNAs, there is a 19-nucleotide segment encompassing the entire T psi C stem and loop region, that, except for post-transcriptional modifications, is identical in this spinach chloroplast tRNAThr and in yeast cytoplasmic tRNAThr1A. A most unusual feature of this spinach chloroplast tRNAThr is that it has an A residue at the 5'-end of the anticodon loop, a site that is occupied by a pyrimidine in all other tRNAs sequenced to date. Other than this feature, the tRNAThr contains all of the invariant and semi-invariant residues normally found in tRNAs. This tRNA hybridizes to spinach chloroplast DNA, but does not hybridize to the same region of the spinach chloroplast genome that contains the genes for spinach chloroplast tRNA1Thr or tRNA2Thr. This tRNA therefore appears to be a third isoaccepting species of threonine tRNA encoded by spinach chloroplast DNA.

Macronuclear response of Paramaecium multimicronucleatum to L-lysine.

A strain of Paramaecium multimicronucleatum was exposed to a medium containing L-lysine; the concentrations of the amino acid were 0.1%, 0.5% and 1.0% for different sets of experiments. In these two latter concentrations, the macronucleus of the ciliate broke down into innumerable small fragments, the microspheres. The micro-nuclei remained inert. The microspheres left the body of paramaecium as cell-free, self-duplicating entities constituted of DNA and RNA and enveloped by a protein coat. They had no nuclear membrane and they resembled the prokaryotes. Grown in culture medium with 0.1% horse serum, the microspheres transformed into small amoebae having typical eukaryotic features. These amoebae maintained a typical cyst-trophic cycle during the successive sub-cultures; they had no similarity with the paramaecia.

Dinoflagellate evolution: speculation and evidence.

Nuclear features of dinoflagellates that were used originally to support the Mesocaryota concept are reviewed. The fibrillar diameter of dinoflagellage chromatin, low level of chromosomal basic proteins, membrane attachment of chromosomes and swirl pattern observed in sectioned chromosomes are features that support a prokaryotic affinity. The presence of repeated and highly complex DNA, a S-phase of DNA synthesis in the cell cycle, presence of basic proteins, and the reinterpretation of extranuclear microtubules as a spindle support the contention that dinoflagellates are eukaryotes. This combination of prokaryotic and eukaryotic features suggests that dionflagellates are a geologically old group and that perhaps they diverged from the higher eukaryotic lineage before evolution of eukaryotic chromatin but after the evolution of repeated DNA. The 2 patterns of carotenoid composition exemplified by the presence of peridinin or fucoxanthin suggest separate origins of dinoflagellate plastids, perhaps by prokaryotic and eukaryotic capture. It is suggested that the species possessing fucoxanthin obtained their plastids by capture of photosynthetic eukaryotes.