PubMed Health⌕ Search

SEARCH · PubMed Health

Results for “Genome Components”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 55 records · Page 3Linked to original sources

Comparative physical mapping links conservation of microsynteny to chromosome structure and recombination in grasses.

Nearly finished sequences for model organisms provide a foundation from which to explore genomic diversity among other taxonomic groups. We explore genome-wide microsynteny patterns between the rice sequence and two sorghum physical maps that integrate genetic markers, bacterial artificial chromosome (BAC) fingerprints, and BAC hybridization data. The sorghum maps largely tile a genomic component containing 41% of BACs but 80% of single-copy genes that shows conserved microsynteny with rice and partially tile a nonsyntenic component containing 46% of BACs but only 13% of single-copy genes. The remaining BACs are centromeric (4%) or unassigned (8%). The two genomic components correspond to cytologically discernible "euchromatin" and "heterochromatin." Gene and repetitive DNA distributions support this classification. Greater microcolinearity in recombinogenic (euchromatic) than nonrecombinogenic (heterochromatic) regions is consistent with the hypothesis that genomic rearrangements are usually deleterious, thus more likely to persist in nonrecombinogenic regions by virtue of Muller's ratchet. Interchromosomal centromeric rearrangements may have fostered diploidization of a polyploid cereal progenitor. Model plant sequences better guide studies of related genomes in recombinogenic than nonrecombinogenic regions. Bridging of 35 physical gaps in the rice sequence by sorghum BAC contigs illustrates reciprocal benefits of comparative approaches that extend at least across the cereals and perhaps beyond.

Base Sequence↗

On the roles of repetitive DNA elements in the context of a unified genomic-epigenetic system.

Repetitive DNA sequences comprise a substantial portion of most eukaryotic and some prokaryotic chromosomes. Despite nearly forty years of research, the functions of various sequence families as a whole and their monomer units remain largely unknown. The inability to map specific functional roles onto many repetitive DNA elements (REs), coupled with the taxon-specificity of sequence families, have led many to speculate that these genomic components are "selfish" replicators generating genomic "junk." The purpose of this paper is to critically examine the selfishness, evolutionary effects, and functionality of REs. First, a brief overview of the range of ideas pertaining to RE function is presented. Second, the argument is presented that the selfish DNA "hypothesis" is actually a narrative scheme, that it serves to protect neo-Darwinian assumptions from criticism, and that this story is untestable and therefore not a hypothesis. Third, attempts to synthesize the selfish DNA concept with complex systems models of the genome and RE functionality are critiqued. Fourth, the supposed connection between RE-induced mutations and macroevolutionary events are stated to be at variance with empirical evidence and theoretical considerations. Hypotheses that base phylogenetic transitions in repetitive sequence changes thus remain speculative. Fifth and finally, the case is made for viewing REs as integrally functional components of chromosomes, genomes, and cells. It is argued throughout that a new conceptual framework is needed for understanding the roles of repetitive DNA in genomic/epigenetic systems, and that neo-Darwinian "narratives" have been the primary obstacle to elucidating the effects of these enigmatic components of chromosomes.

Animals↗

Intersubgenomic heterosis in seed yield potential observed in a new type of Brassica napus introgressed with partial Brassica rapa genome.

This paper reports the observation on the intersubgenomic heterosis for seed yield among hybrids between natural Brassica napus (A(n)A(n)C(n)C(n)) and a new type of B. napus with introgressions of genomic components of Brassica rapa (A(r)A(r)). This B. napus was selected from the progeny of B. napus x B. rapa and (B. napus x B. rapa) x B. rapa based on extensive phenotypic and cytological observation. Among the 129 studied partial intersubgenomic hybrids, which were obtained by randomly crossing 13 lines of the new type of B. napus in F(3) or BC(1)F(3) to 27 cultivars of B. napus from different regions as tester lines, about 90% of combinations exceeded the yield of their respective tester lines, whereas about 75% and 25% of combinations surpassed two elite Chinese cultivars, respectively. This strong heterosis was further confirmed by reevaluating 2 out of the 129 combinations in a successive year and by surveying hybrids between 20 lines of the new type of B. napus in BC(1)F(5) and its parental B. napus in two locations. Some DNA segments from B. rapa were identified with significant effects on seed yield and yield components of the new type of B. napus in BC(1)F(5) and intersubgenomic hybrids in positive or negative direction. It seems that the genomic components introgressed from B. rapa contributed to improvement of seed yield of rapeseed.

Analysis of Variance↗

Complementable and noncomplementable host adaptation defects in bipartite geminiviruses.

Members of the geminvirus group of plant viruses collectively infect a broad spectrum of species. Individual viruses which are genetically very similar may nevertheless have distinct host ranges. Two such geminiviruses are tomato golden mosaic virus (TGMV) and been golden mosaic virus (BGMV), for which common hosts have not previously been reported. Each virus has two genome components, designated A and B. The A component is capable of autonomous replication and encapsidation, whereas the B component provides viral functions required for the spread of infection in plants. To investigate the basis for the distinctive host ranges of BGMV and TGMV, we have introduced plasmids containing cloned viral genome components into Nicotiana benthamiana, a good host for TGMV, and bean, Phaseolus vulgaris, a good host for BGMV. We found that TGMV has a low specific infectivity for bean and is virulent, whereas BGMV has a high specific infectivity for N. benthamiana, but infections are asymptomatic and viral DNA accumulation is low. To investigate which viral functions were defective in the poor host in each case, we attempted to complement them by co-inoculation with the well-adapted virus. After inoculation of beans with both viruses, only BGMV was detected. Thus, TGMV exhibits a noncomplementable host adaptation defect in beans. This suggests that the defect has a cis-acting or virus-specific trans-acting genetic basis. In contrast, the BGMV phenotype of low DNA accumulation in N. benthamiana was partially complemented by TGMV A alone and complemented further by the complete TGMV genome. This suggests that a virus nonspecific, trans-acting factor encoded by the BGMV A component is poorly adapted to N. benthamiana. The results of this study indicate that bipartite geminivirus host range may be limited by defective virus-host interactions of more than one kind.

DNA, Viral↗

Pseudorecombination and complementation between potato yellow mosaic geminivirus and tomato golden mosaic geminivirus.

Pseudorecombinants made by exchanging the cloned, infectious genome components (DNAs A and B) of potato yellow mosaic geminivirus (PYMV) and the common strain (cs) of tomato golden mosaic geminivirus (csTGMV) are not infectious in their common host Nicotiana benthamiana. In an N. benthamiana leaf disc assay neither PYMV DNA A nor TGMV DNA A transreplicated each other's DNA B component. The ability of PYMV and TGMV to mediate the systemic movement of each other's DNA A was investigated following coinoculation of N. benthamiana with both genome components of one virus (the helper virus) and DNA A of the other virus (the dependent virus). Movement of the dependent virus DNA A in both cases illustrates interchangeability between the DNA B-encoded movement proteins of New World geminiviruses which infect solanaceous hosts. We have studied this genetic interchangeability further in separate co-agroinoculation experiments with N. benthamiana plants using TGMV DNA A to complement mutations in PYMV open reading frame (ORF) AC2, which encodes a protein that trans-activates the expression of virion sense promoters, and in PYMV ORF AC3, which specifies a protein that enhances viral DNA replication. TGMV DNA A complemented a PYMV AC2 mutant and restored its infectivity and it also complemented a PYMV AC3 mutant and restored the reduced DNA phenotype.

Base Sequence↗

bioWidgets: data interaction components for genomics.

MOTIVATION: The presentation of genomics data in a perspicuous visual format is critical for its rapid interpretation and validation. Relatively few public database developers have the resources to implement sophisticated front-end user interfaces themselves. Accordingly, these developers would benefit from a reusable toolkit of user interface and data visualization components. RESULTS: We have designed the bioWidget toolkit as a set of JavaBean components. It includes a wide array of user interface components and defines an architecture for assembling applications. The toolkit is founded on established software engineering design patterns and principles, including componentry, Model-View-Controller, factored models and schema neutrality. As a proof of concept, we have used the bioWidget toolkit to create three extendible applications: AnnotView, BlastView and AlignView.

Amino Acid Sequence↗

Kinetics of tomato golden mosaic virus DNA replication and coat protein promoter activity in Nicotiana tabacum protoplasts.

We have analyzed the replication kinetics of the DNA A and DNA B genome components of the geminivirus tomato golden mosaic virus (TGMV) in protoplasts derived from Nicotiana tabacum suspension culture. In addition, the kinetics of TGMV coat protein promoter activity, as measured by expression of a beta-glucuronidase (GUS) reporter, have been examined. In our protoplast system, double-stranded DNA forms of both viral genome components appeared by 18 hr post-transfection, while single-stranded DNA accumulated to detectable levels after 18-24 hr. Expression of GUS from the TGMV coat protein promoter did not require viral DNA replication, nor was it dependent on expression of AL1, the only viral gene necessary for DNA replication. However, maximal expression was achieved following AL1-mediated replication of DNA A. GUS activity from replicating templates exceeded that from nonreplicating templates by 60- to 90-fold. Expression of the GUS reporter gene from nonreplicating viral DNA templates was similar to GUS expression from the 35S promoter of cauliflower mosaic virus in N. tabacum protoplasts.

Blotting, Southern↗

Size reversion of African cassava mosaic virus coat protein gene deletion mutants during infection of Nicotiana benthamiana.

Mutants of African cassava mosaic virus containing extensive deletions across the coat protein gene that remove up to one-third of the genomic component have been constructed and shown to be infectious when mechanically inoculated onto Nicotiana benthamiana by leaf abrasion. Using N. tabacum protoplasts we demonstrate that mutant pCLV.CP delta 11, containing a 712 bp deletion, is competent for replication in its deleted form. However, systemic spread of pCLV. CP delta 11 and other deletion mutants is associated with reversion of DNA 1 to a size comparable to that of the native genomic component. This contrasts with the behaviour of coat protein mutants of the closely related geminivirus tomato golden mosaic virus which maintain their deletions during spread. Appraisal of the different inoculation procedures used to introduce the mutants into plants suggests the imposition of a stringent size requirement for localized cell-to-cell spread which is relaxed for long distance spread through the vascular system.

Base Sequence↗

DNA and RNA polymerase activities of nuclei and hypotonic extracts of nuclei isolated from tomato golden mosaic virus infected tobacco leaves.

Nuclei and hypotonically leached extracts of nuclei prepared from tomato golden mosaic virus (TGMV)-infected Nicotiana benthamiana leaves have been used in in vitro DNA and RNA polymerisation reactions. The synthesis of virus-specific DNA was resistant to aphidicolin, sensitive to N-ethylmaleimide and dideoxy TTP, and stimulated by KC1 and ATP. Variably virion (+) and complementary (-) strand DNA of both the A and B genomic components were synthesised. Virus-specific RNA was synthesised in reactions which were initiated prior to nuclei isolation and leaching. From inhibitor studies and salt requirements RNA synthesis appeared to be catalysed by a DNA-dependent RNA polymerase type II enzyme. Both components of the TGMV genome were transcribed in a bidirectional fashion with a prevalence in some experiments of transcripts derived from DNA component A.

Cell Fractionation↗

The P gene product of hepatitis B virus is required as a structural component for genomic RNA encapsidation.

Encapsidation of the pregenomic RNA into nucleocapsids is a selective process which depends on specific RNA-protein interactions. The signal involved in the packaging of the hepatitis B virus (HBV) RNA pregenome was recently defined as a short sequence located near the 5' end of that molecule (Junker-Niepmann et al., EMBO J., in press), but it remained an open question which viral proteins are required. Using a genetic approach, we analyzed whether proteins derived from the HBV P gene play an important role in pregenome encapsidation. The results obtained with point mutations, deletions, and insertions scattered throughout the P gene clearly demonstrate that (i) a P gene product containing all functional domains is required both for the encapsidation of HBV pregenomic RNA and for packaging of nonviral RNAs fused to the HBV encapsidation signal, (ii) known enzymatic activities are not involved in the packaging reaction, suggesting that P protein is required as a structural component, and (iii) P protein acts primarily in cis, i.e., pregenomic RNAs from which P protein is synthesized are preferentially encapsidated.

Base Sequence↗

Whole-genome variance components linkage analysis using single-nucleotide polymorphisms versus microsatellites on quantitative traits of derived phenotypes from factor analysis of electroencephalogram waves.

Alcohol dependence is a serious public health problem. We studied data from families participating in the Collaborative Study on the Genetics of Alcoholism (COGA) and made available to participants in the Genetic Analysis Workshop 14 (GAW14) in order to search for genes predisposing to alcohol dependence. Using factor analysis, we identified four factors (F1, F2, F3, F4) related to the electroencephalogram traits. We conducted variance components linkage analysis with each of the factors. Our results using the Affymetrix single-nucleotide polymorphism dataset showed significant evidence for a novel linkage of F3 (factor comprised of the three midline channel EEG measures from the target case of the Visual Oddball experiment ttdt2, 3, 4) to chromosome 18 (LOD = 3.45). This finding was confirmed by analyses of the microsatellite data (LOD = 2.73) and Illumina SNP data (LOD = 3.30). We also demonstrated that, in a sample like the COGA data, a dense single-nucleotide polymorphism map provides better linkage signals than low-resolution microsatellite map with quantitative traits.

Chromosome Mapping↗

DNA turnover and the molecular clock.

Many detailed studies on the mechanisms by which different components of eukaryotic nuclear genomes have diverged reveal that the majority of sequences are seemingly not passively accumulating base substitutions in a clocklike manner solely determined by laws of diffusion at the population level. It appears that variation in the rates, units, biases, and gradients of several DNA turnover mechanisms are contributing to the course of DNA divergence. Turnover mechanisms have the potential to retard, maintain, or accelerate the rate of DNA differentiation between populations. Furthermore, examples are known of coding and noncoding DNA subject to the simultaneous operation of several turnover mechanisms leading to complex patterns of fine-scale restructuring and divergence, generally uninterpretable using selection and/or neutral drift arguments in isolation. Constancy in the rate of divergence, where observed over defined periods of time, could be a reflection of constancy in the rates and units of turnover. However, a consideration of the generally large disparity between rates of turnover and mutation reveals that DNA clocks, which would be independently driven by turnover in separate genomic components, would tend to be episodic. The utility of any given DNA sequence for measuring time and species relationships, like individual proteins, is proportional to the extent to which all contributing forces to the evolution of the sequence, internal and external, are understood.

Biological Evolution↗

A cluster of dispensable genes within the human cytomegalovirus genome short component: IRS1, US1 through US5, and the US6 family.

By insertional mutagenesis, human cytomegalovirus recombinants deleted of each of the US6 glycoprotein family genes were isolated. A recombinant lacking IRS1, US1 through US5, and most of the US6 family was also isolated. The growth kinetics of these mutants were similar to that of the wild type. A dispensable cluster of genes was identified.

Cells, Cultured↗

Nucleic acids in mummified plant seeds: biochemistry and molecular genetics of pre-Columbian maize.

Nucleic acids fractions were isolated from pre-Columbian maize seeds and characterized using different approaches such as polyacrylamide gel electrophoresis, anti-DNA antibody binding, HPLC fractionation, molecular hybridization with cloned genes, and DNA amplification by the polymerase chain reaction. The nucleic acids were found to be very depolymerized (less than or equal to 140 base pairs in length) and composed mainly of ribosomal RNA. Despite the very low amount and degree of polymerization of seed DNA, specific maize nuclear Mu1, Mu4, Mu8 and, possibly, Mu5 element components could be detected, thanks to the use of amplification systems as short as 90 bp. The results suggest that evaluation of the relative proportions of Mu-type element components and, possibly, other maize genomic components in single mummified kernels, may offer a new key to the study of ancient maize populations.

Base Sequence↗

Molecular biodiversity of cassava begomoviruses in Tanzania: evolution of cassava geminiviruses in Africa and evidence for East Africa being a center of diversity of cassava geminiviruses.

Cassava is infected by numerous geminiviruses in Africa and India that cause devastating losses to poor farmers. We here describe the molecular diversity of seven representative cassava mosaic geminiviruses (CMGs) infecting cassava from multiple locations in Tanzania. We report for the first time the presence of two isolates in East Africa: (EACMCV-[TZ1] and EACMCV-[TZ7]) of the species East African cassava mosaic Cameroon virus, originally described in West Africa. The complete nucleotide sequence of EACMCV-[TZ1] DNA-A and DNA-B components shared a high overall sequence identity to EACMCV-[CM] components (92% and 84%). The EACMCV-[TZ1] and -[TZ7] genomic components have recombinations in the same genome regions reported in EACMCV-[CM], but they also have additional recombinations in both components. Evidence from sequence analysis suggests that the two strains have the same ancient origin and are not recent introductions. EACMCV-[TZ1] occurred widely in the southern part of the country. Four other CMG isolates were identified: two were close to the EACMV-Kenya strain (named EACMV-[KE/TZT] and EACMV-[KE/TZM] with 96% sequence identity); one isolate, TZ10, had 98% homology to EACMV-UG2Svr and was named EACMV-UG2 [TZ10]; and finally one isolate was 95% identical to EACMV-[TZ] and named EACMV-[TZ/YV]. One isolate of African cassava mosaic virus with 97% sequence identity with other isolates of ACMV was named ACMV-[TZ]. It represents the first ACMV isolate from Tanzania to be sequenced. The molecular variability of CMGs was also evaluated using partial B component nucleotide sequences of 13 EACMV isolates from Tanzania. Using the sequences of all CMGs currently available, we have shown the presence of a number of putative recombination fragments that are more prominent in all components of EACMV than in ACMV. This new knowledge about the molecular CMG diversity in East Africa, and in Tanzania in particular, has led us to hypothesize about the probable importance of this part of Africa as a source of diversity and evolutionary change both during the early stages of the relationship between CMGs and cassava and in more recent times. The existence of multiple CMG isolates with high DNA genome diversity in Tanzania and the molecular forces behind this diversity pose a threat to cassava production throughout the African continent.

Base Sequence↗

Characterization of the Nicotiana tabacum L. genome by molecular cytogenetics.

Nicotiana tabacum (2n = 48) is a natural amphidiploid with component genomes S and T. We used non-radioactive in situ hybridization to provide physical chromosome markers for N. tabacum, and to determine the extant species most similar to the S and T genomes. Chromosomes of the S genome hybridized strongly to biotinylated total DNA from N. sylvestris, and showed the same physical localization of a tandemly repeated DNA sequence, HRS 60.1, confirming the close relationship between the S genome and N. sylvestris. Results of dot blot and in situ hybridizations of N. tabacum DNA to biotinylated total genomic DNA from N. tomentosiformis and N. otophora suggested that the T genome may derive from an introgressive hybrid between these two species. Moreover, a comparison of nucleolus-organizing chromosomes revealed that the nucleolus organizer region (NOR) most strongly expressed in N. tabacum had a very similar counterpart in N. otophora. Three different N. tabacum genotypes each had up to 9 homozygous translocations between chromosomes of the S and T genomes. Such translocations, which were either unilateral or reciprocal, demonstrate that intergenomic transfer of DNA has occurred in the amphidiploid, possibly accounting for some results of previous genetic and molecular analyses. Molecular cytogenetics of N. tabacum has identified new chromosome markers, providing a basis for physical gene mapping and showing that the amphidiploid genome has diverged structurally from its ancestral components.

Chromosome Mapping↗

The intergenic region of maize streak virus contains promoter elements involved in rightward transcription of the viral genome.

Maize streak virus (MSV), a geminivirus with a one-component genome, encodes a major coat protein RNA which accumulates in infected plants. Using a maize protoplast cell transient expression system, we have defined and studied the promoter which drives rightward transcription of the RNA encoding the coat protein. We have identified a 122 bp upstream segment that enhances promoter activity and functions as an upstream activating sequence (UAS). The UAS lies in the starting intergenic region of the viral genome and includes a region which is similar in all geminiviruses. The 122 bp UAS activates the MSV core promoter in an orientation, but not position, independent fashion. The MSV promoter UAS is interchangeable with a similar element in the cauliflower mosaic virus (CaMV) 35S RNA core promoter, that is the MSV UAS will activate the CaMV 35S core promoter and vice versa. However, the MSV promoter UAS specifically binds proteins in maize nuclear extracts which appear to differ from those bound by the functionally equivalent region of the CaMV 35S promoter.

Journal Article↗

Development of broadly targeted human endogenous gammaretroviral pol-based real time PCRs Quantitation of RNA expression in human tissues.

Endogenous retroviral sequences (ERVs) are dynamic genomic components with profound influences on gene expression and genomic structure. Their extent of expression is not well known. Several broadly targeted real-time reverse transcription PCR (QPCRs) systems for surveillance of RNA expression of the major groups of human gammaretroviral ERVs were constructed. The highly conserved reverse transcriptase (RT) and integrase (IN) domains of the pol gene were used as targets for the PCRs, which were both probe-based (TaqMan) and probe-less (SYBR Green). Different levels of primer and probe degeneracy, with or without inosine, were tested. Several of the PCRs had sensitivities of a few HERV nucleic acid copies per PCR reaction. Specificities were approximately as expected from the fit of primers and probes. Gammaretroviral HERV RNA expression was studied in different human tissues. Each HERV group had a specific pattern of expression. HERV-E was highly expressed in testis, HERV-I/T in brain and testis, HERV-H in brain and testis, while HERV-W was highly expressed in placenta. Gammaretroviral RNA was not detected in plasma from 50 blood donors in saliva from 20 persons. In conclusion, a set of tools for investigation of gammaretroviral HERV RNA expression was created.

Base Sequence↗