Burnet oration: dendritic cells: multiple subtypes, multiple origins, multiple functions

The dendritic cells (DC) of the mouse are surprisingly heterogeneous by surface phenotype and may be segregated based on expression of CD4 and CD8. These DC subtypes appear to differ in developmental origin and display some differences in biological function, including regulation of the cytokine production of the T cells that they activate. This presentation reviews the attempts of one laboratory to understand this complex DC system.     

The DNA-binding domain of simian virus 40 tumor antigen has multiple functions.

The DNA-binding domain of simian virus 40 tumor antigen has been previously shown to participate in a number of different activities. Besides being involved in binding to sequences at the viral replication origin, this domain appears to be required for nonspecific DNA binding, for structurally distorting origin DNA (melting and untwisting), and possibly for oligomerization of the protein into hexamers and double hexamers. We now provide evidence that it also takes part in unwinding origin DNA sequences, contributes a function specifically related to in vivo DNA replication, and perhaps supports the assembly of the virus or release of the virus from the cell. This 100-amino-acid domain appears to be an excellent model system for studying how a small region of a protein could have a number of distinct activities.     

The multiple evolutionary histories of dioxygen reductases: Implications for the origin and evolution of aerobic respiration

Understanding the origin and evolution of cellular processes is fundamental to understand how biological activity has shaped the history of our planet. Among these, aerobic respiration is probably one of the most debated. We have applied a phylogenomics approach to investigate the origin and evolution of dioxygen reductases (O(2)Red), the key enzymes of aerobic respiratory chains. The distribution and phylogenetic analysis of the four types of O(2)Red (Cyt-bd and the A, B, and C families of heme-copper O(2)Red) from 673 complete bacterial and archaeal genomes show that these enzymes have very different evolutionary histories: Cyt-bd are of bacterial origin and were transferred to a few archaea; C-O(2)Red are of proteobacterial origin and were transferred to a few other bacteria; B-O(2)Red are of archaeal origin and were transferred to a few bacteria; and A-O(2)Red are the most ancient O(2)Red and were already present prior to the divergence of major present-day bacterial and archaeal phyla, thus before the emergence of Cyanobacteria and oxygenic photosynthesis. Implications for the origin and the evolution of aerobic respiration are discussed.     

Polyploid hybrids: multiple origins of a treefrog species

Hyla versicolor, a tetraploid treefrog, is reported to have originated via multiple hybridization events involving three diploid ancestors. Its complex reticulate history provides insights into the roles that polyploidy and hybridization can play in the origin of species.     

Mitochondrial DNA reveals multiple introductions of domestic chicken in East Africa

Chicken were possibly domesticated in South and Southeast Asia. They occur ubiquitously in East Africa where they show extensive phenotypic diversity. They appeared in the region relatively late, with the first undisputed evidence of domestic chicken in Sudan, around ~ 700 BC. We reveal through a detailed analysis of mitochondrial DNA D-loop sequence diversity of 512 domestic village chickens, from four East African countries (Kenya, Ethiopia, Sudan, Uganda), the presence of at least five distinct mitochondrial DNA haplogroups. Phylogeographic analyses and inclusion of reference sequences from Asia allow us to address the origin, ways of introduction and dispersion of each haplogroup. The results indicate a likely Indian subcontinent origin for the commonest haplogroup (D) and a maritime introduction for the next commonest one (A) from Southeast and/or East Asia. Recent introgression of commercial haplotypes into the gene pool of village chickens might explain the rare presence of two haplogroups (B and C) while the origin of the last haplogroup (E) remains unclear being currently observed only outside the African continent in the inland Yunnan Province of China. Our findings not only support ancient historical maritime and terrestrial contacts between Asia and East Africa, but also indicate the presence of large maternal genetic diversity in the region which could potentially support genetic improvement programmes.     

Simian virus 40 DNA replication in vitro: identification of multiple stages of initiation.

A cell-free DNA replication system dependent upon five purified cellular proteins, one crude cellular fraction, and the simian virus 40 (SV40)-encoded large tumor antigen (T antigen) initiated and completed replication of plasmids containing the SV40 origin sequence. DNA synthesis initiated at or near the origin sequence after a time lag of approximately 10 min and then proceeded bidirectionally from the origin to yield covalently closed, monomer daughter molecules. The time lag could be completely eliminated by a preincubation of SV40 ori DNA in the presence of T antigen, a eucaryotic single-stranded DNA-binding protein (replication factor A [RF-A]), and topoisomerases I and II. In contrast, if T antigen and the template DNA were incubated alone, the time lag was only partially decreased. Kinetic analyses of origin recognition by T antigen, origin unwinding, and DNA synthesis suggest that the time lag in replication was due to the formation of a complex between T antigen and DNA called the T complex, followed by formation of a second complex called the unwound complex. Formation of the unwound complex required RF-A. When origin unwinding was coupled to DNA replication by the addition of a partially purified cellular fraction (IIA), DNA synthesis initiated at the ori sequence, but the template DNA was not completely replicated. Complete DNA replication in this system required the proliferating-cell nuclear antigen and another cellular replication factor, RF-C, during the elongation stage. In a less fractionated system, another cellular fraction, SSI, was previously shown to be necessary for reconstitution of DNA replication. The SSI fraction was required in the less purified system to antagonize the inhibitory action of another cellular protein(s). This inhibitor specifically blocked the earliest stage of DNA replication, but not the later stages. The implications of these results for the mechanisms of initiation and elongation of DNA replication are discussed.     

A congruent molecular signature of vicariance across multiple plant lineages

Explaining disjunct distributions, or why closely related organisms are often separated by apparently severe barriers such as oceans or deserts, is a great challenge for historical biogeography. Competing explanations are long-distance dispersal across a barrier, and vicariance, in which disjunct taxa are descended from an ancestral population that was split by formation of the barrier. Vicariance explanations are testable by their prediction that near-simultaneous speciation should have occurred across multiple lineages of organisms between the disjunct areas because the origin of a barrier would potentially disrupt gene flow within multiple species. To date, there have been few studies providing evidence for multiple synchronous ancient divergences across a barrier whose origin coincides with the timing of the speciation events. Here, we use relaxed molecular-clock dating to investigate the timing of south-western (SW) versus south-eastern (SE) divergences in 23 pairs of plant lineages in southern Australia. Sixteen of the divergences correlate with the origin, 13-14 million years (Myr) ago, of the arid treeless Nullarbor Plain. The Nullarbor Plain currently forms a substantial barrier to SW-SE migration but during the last 45Myr this region has experienced multiple episodes of marine inundation and subaerial exposure. Thus, there have been multiple events that could have caused either isolation and speciation, or secondary contact, among the taxa of southern Australia. The strong molecular signal of coincident speciation in many diverse lineages during a short period provides the best evidence to date linking synchronous speciation to an ancient vicariance event.     

Parent-of-origin effects in multiple endocrine neoplasia type 2B.

Multiple endocrine neoplasia type 2B (MEN 2B) is characterized by medullary thyroid carcinoma, pheochromocytomas, mucosal neuromas, ganglioneuromas, and skeletal and ophthalmic abnormalities. It is observed as both inherited and sporadic disease, with an estimated 50% of cases arising de novo. A single point mutation in the catalytic core region of the receptor tyrosine kinase, RET, has been observed in germ-line DNA of MEN 2B patients. We have analyzed 25 cases of de novo disease in order to determine the parental origin of the mutated RET allele. In all cases the new mutation was of paternal origin. We observe a distortion of the sex ratio in both de novo MEN 2B patients and the affected offspring of MEN 2B transmitting males. These results suggests a differential susceptibility of RET to mutation in paternally and maternally derived DNA and a possible role for imprinting of RET during development.     

Large T-antigen mutants define multiple steps in the initiation of simian virus 40 DNA replication.

The biochemical activities of a series of transformation-competent, replication-defective large T-antigen point mutants were examined. The assays employed reflect partial reactions required for the in vitro replication of simian virus 40 (SV40) DNA. Mutants which failed to bind specifically to SV40 origin sequences bound efficiently to single-stranded DNA and exhibited nearly wild-type levels of helicase activity. A mutation at proline 522, however, markedly reduced ATPase, helicase, and origin-specific unwinding activities. This mutant bound specifically to the SV40 origin of replication, but under certain conditions it was defective in binding to both single-stranded DNA and the partial duplex helicase substrate. This suggests that additional determinants outside the amino-terminal-specific DNA-binding domain may be involved in nonspecific binding of T antigen to single-stranded DNA and demonstrates that origin-specific DNA binding can be separated from binding to single-stranded DNA. A mutant containing a lesion at residue 224 retained nearly wild-type levels of helicase activity and recognized SV40 origin sequences, yet it failed to function in an origin-specific unwinding assay. This provides evidence that origin recognition and helicase activities are not sufficient for unwinding to occur. The distribution of mutant phenotypes reflects the complex nature of the initiation reaction and the multiplicity of functions provided by large T antigen.     

Cells with multiple chromosome aberrations in control individuals

Chromosome analysis for asymmetrical chromosome aberrations was performed on 48-h lymphocyte cultures from 12 young trainees who had recently commenced employment, in order to obtain control levels. In two of them, cells were found with multiple breaks and exchanges but when repeated 50 days later no evidence of such cells was found. The possible factors responsible for the origin of these cells and their subsequent disappearance are discussed with the conclusion that a viral etiology seems the most likely.     

The evolutionary history of the white-rayed species of Melampodium (Asteraceae) involved multiple cycles of hybridization and polyploidization

? Premise of the study: Polyploidy plays an important role in race differentiation and eventually speciation. Underlying mechanisms include chromosomal and genomic changes facilitating reproductive isolation and/or stabilization of hybrids. A prerequisite for studying these processes is a sound knowledge on the origin of polyploids. A well-suited group for studying polyploid evolution consists of the three species of Melampodium ser. Leucantha (Asteraceae): M. argophyllum, M. cinereum, and M. leucanthum. ? Methods: The origin of polyploids was inferred using network and tree-based phylogenetic analyses of several plastid and nuclear DNA sequences and of fingerprint data (AFLP). Genome evolution was assessed via genome size measurements, karyotype analysis, and in situ hybridization of ribosomal DNA. ? Key results: Tetraploid cytotypes of the phylogenetically distinct M. cinereum and M. leucanthum had, compared to the diploid cytotypes, doubled genome sizes and no evidence of gross chromosomal rearrangements. Hexaploid M. argophyllum constituted a separate lineage with limited intermixing with the other species, except in analyses from nuclear ITS. Its genome size was lower than expected if M. cinereum and/or M. leucanthum were involved in its origin, and no chromosomal rearrangements were evident. ? Conclusions: Polyploids in M. cinereum and M. leucanthum are of recent autopolyploid origin in line with the lack of significant genomic changes. Hexaploid M. argophyllum also appears to be of autopolyploid origin against the previous hypothesis of an allopolyploid origin involving the other two species, but some gene flow with the other species in early phases of differentiation cannot be excluded.     

A perivascular origin for mesenchymal stem cells in multiple human organs

Mesenchymal stem cells (MSCs), the archetypal multipotent progenitor cells derived in cultures of developed organs, are of unknown identity and native distribution. We have prospectively identified perivascular cells, principally pericytes, in multiple human organs including skeletal muscle, pancreas, adipose tissue, and placenta, on CD146, NG2, and PDGF-Rbeta expression and absence of hematopoietic, endothelial, and myogenic cell markers. Perivascular cells purified from skeletal muscle or nonmuscle tissues were myogenic in culture and in vivo. Irrespective of their tissue origin, long-term cultured perivascular cells retained myogenicity; exhibited at the clonal level osteogenic, chondrogenic, and adipogenic potentials; expressed MSC markers; and migrated in a culture model of chemotaxis. Expression of MSC markers was also detected at the surface of native, noncultured perivascular cells. Thus, blood vessel walls harbor a reserve of progenitor cells that may be integral to the origin of the elusive MSCs and other related adult stem cells.     

DNA replication initiates non-randomly at multiple sites near the c-myc gene in HeLa cells.

The origin of replication of the c-myc gene in HeLa cells was previously identified at low resolution within 3.5 kb 5' to the P1 promoter, based on replication fork polarity and the location of DNA nascent strands. To define the initiation events in the c-myc origin at higher resolution the template bias of nascent DNAs in a 12 kb c-myc domain has been analyzed by hybridization to strand specific probes. Strong switches in the asymmetry of nascent strand template preference confirm that replication initiates non-randomly at multiple sites within 2.4 kb 5' to the c-myc P1 promoter, and at other sites over a region of 12 kb or more. The strongest template biases occur in the 2.4 kb region 5' of the P1 promoter, shown earlier to contain sequences which allow the autonomous semiconservative replication of c-myc plasmids. An asymmetric pyrimidine heptanucleotide consensus sequence has been identified which occurs 12 times in the c-myc origin zone, and whose polarity exactly correlates with the polarity of nascent strand synthesis.     

An exploratory analysis of multiple mutation spectra

The last decade has witnessed a remarkable increase in the number of mutations identified both in human disease-related genes and mutation reporter genes including those in mammalian cells and transgenic animals. This has led to the curation of a number of computerised databases, which make mutation data freely available for analysis. A primary interest of both the clinical researcher and the genetic toxicologist is determination of location and types of mutation within a gene of interest. Collections of mutation data observed for a disease-related gene or, for a gene exposed to a particular chemical, permits discovery of regions of sequence along the gene prone to mutagenesis and may provide clues to the origin of a mutation. The principal tool for visualising the distribution pattern of mutant data along a gene is the mutation spectrum: the distribution and frequency of mutations along a nucleotide sequence. In genetic toxicology, the current wealth of mutation data available allows us to construct many mutation spectra of interest to investigate the mutagenic mechanisms and mutational sites for one or a group of mutagens. Using the multivariate statistical methods principal components analysis (PCA) and cluster analysis (CA) we have tested the ability of these methods to establish the underlying patterns within and between 60 UV-induced, mitomycin C-induced and spontaneous mutations in the supF gene. The spectra were derived from human, monkey and mouse cells including both repair efficient and repair deficient cell lines. We demonstrate and support the successful application of multivariate statistical methods for exploring large sets of mutation spectra to reveal underlying patterns, groupings and similarities. The methods clearly demonstrate how different patterns of spontaneous and UV-induced supF mutation spectra can result from variation in plasmid, culture medium, species origin of cell line and whether mutations arose in vivo or in vitro.     

Fossils and phylogenies: integrating multiple lines of evidence to investigate the origin of early major metazoan lineages

Understanding the nature and timing of metazoan origins is oneof the most important, yet elusive, questions in evolutionarybiology. Fossil data provide the most tangible evidence forthe origin of early animal lineages, although additional evidencefrom molecular phylogenetics, molecular clock studies, and developmenthas contributed to our current understanding. We review severallines of evidence to explore the nature and timing of earlymetazoan evolution and discuss how these data, when consideredtogether, provide a more cohesive picture of the origin of animaldiversity. We discuss how trace fossils and biomarkers providecompelling evidence for the origins of Bilateria and siliceoussponges. Using a molecular phylogenetic framework for metazoans,we discuss how fossils can be used to date the origin of clades.We use these fossil dates to perform a relaxed molecular clockanalysis for estimating dates of nodes when no fossils are available.We also discuss current data from developmental biology thatsuggest that early metazoans possessed a sophisticated moleculartoolkit for building complex body plans. We conclude that thebest evidence for the origin of major metazoan lineages liesin the careful interpretation of the fossil record and thatthese data, when considered with phylogenetic and developmentalevidence, support the notion that the Cambrian radiation isa real phenomenon that marks a critically important time inthe history of life.     

Triploid-diploid mosaicism in the lymphocytes of a liveborn child with multiple malformations

Summary A new case of triploid-diploid mosaicism in the lymphocytes of a newborn with multiple malformations is reported, and the origin of the mosaicism is briefly discussed.N.I.H. International Post-doctoral Fellow.Bevoegd Navorser N.F.W.O.     

Microsatellite markers reveal multiple origins for Italian weedy rice

Weedy rice (Oryza sativa L.) is one of the major issues of rice cultivation worldwide. In Italy, it infests about 70% of the total rice area. Different Weedy Rice populations can be distinguished based on variable morphological and physiological traits; however, little is known about genetic differentiation and origin of Italian weedy rice populations. The objective of this study was to genetically and morphologically characterize and compare different Italian weedy rice populations selected on the basis of different phenotypes. The main Italian rice territory was divided into 10 geographical areas in which 40 weedy rice populations were collected and grouped according to the awn traits. All the individuals of the populations were morphologically characterized according to plant and seed traits. Genetic characterization was performed using 19 SSR markers on all the collected accessions, and several rice cultivars, including some very old (late 19th century), nowadays are no longer cultivated. ANOVA showed that morphological plant and seed traits were significantly affected by the collection area and awnedness group. The importance of the awn morphology was also reflected in the Bayesian clustering where, despite a relatively low genetic diversity, the clusters displayed different awn types. An UPGMA dendrogram confirmed the clusters detected in STRUCTURE analysis and also revealed a grouping of certain old cultivars with the weedy rice, suggesting a common origin.