Chromosomal rearrangements in rock wallabies, Petrogale (Marsupialia: Macropodidae). VIII. An investigation of the nonrandom nature of karyotypic change

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 524-534 ◽  
Author(s):  
M. D. B. Eldridge ◽  
P. G. Johnston
Keyword(s):  
Mitomycin C ◽  
Chromosome Evolution ◽  
Hot Spot ◽  
Chromosomal Rearrangements ◽  
Mitotic Chromosomes ◽  
Chromosome 5 ◽  
Chromosome 10 ◽  
Cultured Fibroblasts ◽  
Chromosome Breaks

Although the nonrandom nature of many chromosome breaks is well known, few studies have investigated the potential significance of this breakage specificity in chromosome evolution. The macropod genus Petrogale is an ideal group in which to investigate this phenomenon, since it comprises a large number of chromosomal forms, many of which appear to have differentiated relatively recently. By exposing Petrogale cells to mutagenic agents it should be possible to compare the distribution and abundance of induced breaks with those that are known to have occurred in vivo during the chromosomal differentiation of the genus. In this study, breaks were induced in mitotic chromosomes from P. assimilis and P. mareeba by exposing synchronized cultured fibroblasts to low doses of gamma radiation. The results were remarkably similar for both species and the distribution of breaks among the chromosomes appeared to be nonrandom. It was found that chromosomes 5, 6, and (possibly) 10 had a substantially higher rate of breakage than expected. These are also the chromosomes that occur disproportionately among the rearrangements identified in Petrogale. While the distribution of breaks along the chromosome appeared uniform or normal for most chromosomes, a putative "hot spot" was identified near the centromere in chromosome 5 of P. mareeba and in a homologous position near the telomere of chromosome 5 in P. assimilis. In a further experiment, a 1- to 2-h pulse of mitomycin C was used to induce centric fusions in cultured fibroblasts of P. penicillata (2n = 22); 2408 cells were examined and 112 fusions were identified. While it was found that all chromosomes participated in forming fusions, chromosome 10 was found to be most frequently involved, being present in 28.6% of the identifed fusions. This frequency is far greater than would be expected if fusions were to occur at random (10%). It is significant then that chromosome 10 has been involved in five of the eight centric fusions that have been identified in Petrogale and that it is also the chromosome that has been most frequently rearranged in Petrogale. These results suggest that features of the karyotype may influence the distribution and frequency of chromosome breaks and therefore the rate and nature of chromosome evolution.Key words: Petrogale, chromosome evolution, irradiation, induced rearrangements, mitomycin C.

Chromosomal rearrangements in rock wallabies, Petrogale (Marsupialia: Macropodidae). III. G-banding analysis of Petrogale inornata and P. penicillata

Genome ◽  
1990 ◽  
Vol 33 (6) ◽  
pp. 798-802 ◽  
Author(s):  
M. D. B. Eldridge ◽  
R. L. Close ◽  
P. G. Johnston
Keyword(s):  
Acrocentric Chromosome ◽  
Chromosomal Rearrangements ◽  
Paracentric Inversion ◽  
Chromosome 3 ◽  
Chromosome 4 ◽  
Chromosome 5 ◽  
Cultured Fibroblasts ◽  
Submetacentric Chromosome ◽  
Ancestral Chromosome ◽  
Banding Analysis

The karyotypes of Petrogale inornata and the two currently recognised races of Petrogale penicillata were examined using G-banding from cultured fibroblasts. Petrogale inornata (2n = 22) was found to retain plesiomorphic chromosomes 3 and 4 but possessed an apomorphic inverted chromosome 5 (5i). This 5i appears identical with the 5i found in two other Queensland taxa, Petrogale assimilis and Petrogale godmani, and can be derived from the ancestral chromosome 5 by an extensive paracentric inversion or a centromeric transposition. Petrogale penicillata penicillata (2n = 22) and Petrogale penicillata herberti (2n = 22) both possess the synapomorphic acrocentric chromosome 3, which appears to differ from the plesiomorphic 3 by a small centromeric transposition. Petrogale p. penicillata was also found to be characterised by an apomorphic acrocentric chromosome 4, while P. p. herberti was characterised by an autapomorphic submetacentric chromosome 4. Both apomorphic chromosomes 4 can be related to the plesiomorphic chromosome 4 by centromeric transpositions. Thus although P. inornata is chromosomally distinct it is more closely related to other north Queensland taxa than it is to either P. p. penicillata or P. p. herberti.Key words: chromosomal rearrangements, G-banding, Marsupialia, Petrogale, Macropodidae.

Conversion of mitomycin C to 2,7-diaminomitosene and 10-decarbamoyl 2,7-diaminomitosene in tumour tissue in vivo

1995 ◽  
Vol 35 (4) ◽  
pp. 318-322 ◽  
Author(s):  
Linda Chirrey ◽  
Jeffrey Cummings ◽  
Gavin W. Halbert ◽  
John F. Smyth
Keyword(s):  
Mitomycin C ◽  
Tumour Tissue

Effects of truncated neurofilament proteins on the endogenous intermediate filaments in transfected fibroblasts

1991 ◽  
Vol 99 (2) ◽  
pp. 335-350 ◽  
Author(s):  
S.S. Chin ◽  
P. Macioce ◽  
R.K. Liem
Keyword(s):  
Intermediate Filament ◽  
Dominant Negative ◽  
Deletion Mutants ◽  
Tail Domain ◽  
Cultured Fibroblasts ◽  
Mutant Proteins ◽  
Amino Terminal ◽  
Novel Approach ◽  
Carboxyl Terminal

The expression and assembly characteristics of carboxyl- and amino-terminal deletion mutants of rat neurofilament low Mr (NF-L) and neurofilament middle Mr (NF-M) proteins were examined by transient transfection of cultured fibroblasts. Deletion of the carboxyl-terminal tail domain of either protein indicated that this region was not absolutely essential for co-assembly into the endogenous vimentin cytoskeleton. However, deletion into the alpha-helical rod domain resulted in an inability of the mutant proteins to co-assemble with vimentin into filamentous structures. Instead, the mutant proteins appeared to be assembled into unusual tubular-vesicular structures. Additionally, these latter deletions appeared to act as dominant negative mutants which induced the collapse of the endogenous vimentin cytoskeleton as well as the constitutively expressed NF-H and NF-M cytoskeletons in stably transfected cell lines. Thus, an intact alpha-helical rod domain was essential for normal IF co-assembly whereas carboxyl-terminal deletions into this region resulted in dramatic alterations of the existing type III and IV intermediate filament cytoskeletons in vivo. Deletions from the amino-terminal end into the alpha-helical rod region gave different results. With these deletions, the transfected protein was not co-assembled into filaments and the endogenous vimentin IF network was not disrupted, indicating that these deletion mutants are recessive. The dominant negative mutants may provide a novel approach to studying intermediate filament function within living cells.

An N-Ethyl-N-Nitrosourea Mutagenesis Screen for Epigenetic Mutations in the Mouse

Genetics ◽  
2003 ◽  
Vol 164 (4) ◽  
pp. 1481-1494
Author(s):  
Ivona Percec ◽  
Joanne L Thorvaldsen ◽  
Robert M Plenge ◽  
Christopher J Krapp ◽  
Joseph H Nadeau ◽  
...  
Keyword(s):  
Genomic Imprinting ◽  
X Chromosome ◽  
X Inactivation ◽  
Monoallelic Expression ◽  
Chromosome 15 ◽  
Chromosome 5 ◽  
Chromosome 10 ◽  
Mutagenesis Screen ◽  
Distal Chromosome ◽  
Males And Females

Abstract The mammalian epigenetic phenomena of X inactivation and genomic imprinting are incompletely understood. X inactivation equalizes X-linked expression between males and females by silencing genes on one X chromosome during female embryogenesis. Genomic imprinting functionally distinguishes the parental genomes, resulting in parent-specific monoallelic expression of particular genes. N-ethyl-N-nitrosourea (ENU) mutagenesis was used in the mouse to screen for mutations in novel factors involved in X inactivation. Previously, we reported mutant pedigrees identified through this screen that segregate aberrant X-inactivation phenotypes and we mapped the mutation in one pedigree to chromosome 15. We now have mapped two additional mutations to the distal chromosome 5 and the proximal chromosome 10 in a second pedigree and show that each of the mutations is sufficient to induce the mutant phenotype. We further show that the roles of these factors are specific to embryonic X inactivation as neither genomic imprinting of multiple genes nor imprinted X inactivation is perturbed. Finally, we used mice bearing selected X-linked alleles that regulate X chromosome choice to demonstrate that the phenotypes of all three mutations are consistent with models in which the mutations have affected molecules involved specifically in the choice or the initiation of X inactivation.

scholarly journals Topoisomerase I involvement in illegitimate recombination in Saccharomyces cerevisiae.

1996 ◽  
Vol 16 (4) ◽  
pp. 1805-1812 ◽  
Author(s):  
J Zhu ◽  
R H Schiestl
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Chromosome Aberrations ◽  
Topoisomerase I ◽  
Wild Type Strain ◽  
Hot Spot ◽  
Illegitimate Recombination ◽  
Wild Type ◽  
Yeast Saccharomyces Cerevisiae

Chromosome aberrations may cause cancer and many heritable diseases. Topoisomerase I has been suspected of causing chromosome aberrations by mediating illegitimate recombination. The effects of deletion and of overexpression of the topoisomerase I gene on illegitimate recombination in the yeast Saccharomyces cerevisiae have been studied. Yeast transformations were carried out with DNA fragments that did not have any homology to the genomic DNA. The frequency of illegitimate integration was 6- to 12-fold increased in a strain overexpressing topoisomerase I compared with that in isogenic control strains. Hot spot sequences [(G/C)(A/T)T] for illegitimate integration target sites accounted for the majority of the additional events after overexpression of topoisomerase I. These hot spot sequences correspond to sequences previously identified in vitro as topoisomerase I preferred cleavage sequences in other organisms. Furthermore, such hot spot sequences were found in 44% of the integration events present in the TOP1 wild-type strain and at a significantly lower frequency in the top1delta strain. Our results provide in vivo evidence that a general eukaryotic topoisomerase I enzyme nicks DNA and ligates nonhomologous ends, leading to illegitimate recombination.

scholarly journals Triple-Helical DNA in Drosophila Heterochromatin

Cells ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 227 ◽  
Author(s):  
Eduardo Gorab
Keyword(s):  
Nucleic Acids ◽  
Dna Sequences ◽  
Detection Methods ◽  
Triplex Dna ◽  
Thiazole Orange ◽  
Chromosomal Dna ◽  
Mitotic Chromosomes ◽  
Satellite Repeats

Polynucleotide chains obeying Watson-Crick pairing are apt to form non-canonical complexes such as triple-helical nucleic acids. From early characterization in vitro, their occurrence in vivo has been strengthened by increasing evidence, although most remain circumstantial particularly for triplex DNA. Here, different approaches were employed to specify triple-stranded DNA sequences in the Drosophila melanogaster chromosomes. Antibodies to triplex nucleic acids, previously characterized, bind to centromeric regions of mitotic chromosomes and also to the polytene section 59E of mutant strains carrying the brown dominant allele, indicating that AAGAG tandem satellite repeats are triplex-forming sequences. The satellite probe hybridized to AAGAG-containing regions omitting chromosomal DNA denaturation, as expected, for the intra-molecular triplex DNA formation model in which single-stranded DNA coexists with triplexes. In addition, Thiazole Orange, previously described as capable of reproducing results obtained by antibodies to triple-helical DNA, binds to AAGAG repeats in situ thus validating both detection methods. Unusual phenotype and nuclear structure exhibited by Drosophila correlate with the non-canonical conformation of tandem satellite arrays. From the approaches that lead to the identification of triple-helical DNA in chromosomes, facilities particularly provided by Thiazole Orange use may broaden the investigation on the occurrence of triplex DNA in eukaryotic genomes.

scholarly journals The HROC-Xenobank—A High Quality Assured PDX Biobank of >100 Individual Colorectal Cancer Models

Cancers ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 5882
Author(s):  
Stephanie Matschos ◽  
Florian Bürtin ◽  
Said Kdimati ◽  
Mandy Radefeldt ◽  
Susann Krake ◽  
...  
Keyword(s):  
Gene Expression Analysis ◽  
Tumor Tissue ◽  
Hot Spot ◽  
In Vivo Studies ◽  
Cooperative Research ◽  
High Quality ◽  
Current Time ◽  
Cancer Models ◽  
Low Passage

Based on our research group’s large biobank of colorectal cancers (CRC), we here describe the ongoing activity of establishing a high quality assured PDX biobank for more than 100 individual CRC cases. This includes sufficient numbers of vitally frozen (n > 30 aliquots) and snap frozen (n > 5) backups, “ready to use”. Additionally, PDX tumor pieces were paraffin embedded. At the current time, we have completed 125 cases. This resource allows histopathological examinations, molecular characterizations, and gene expression analysis. Due to its size, different issues of interest can be addressed. Most importantly, the application of low-passage, cryopreserved, and well-characterized PDX for in vivo studies guarantees the reliability of results due to the largely preserved tumor microenvironment. All cases described were molecularly subtyped and genetic identity, in comparison to the original tumor tissue, was confirmed by fingerprint analysis. The latter excludes ambiguity errors between the PDX and the original patient tumor. A cancer hot spot mutation analysis was performed for n = 113 of the 125 cases entities. All relevant CRC molecular subtypes identified so far are represented in the Hansestadt Rostock CRC (HROC)-Xenobank. Notably, all models are available for cooperative research approaches.

scholarly journals Rapid in vivo development of resistance to daptomycin in vancomycin-resistant Enterococcus faecium due to genomic rearrangements

2021 ◽  
Author(s):  
Sarah Mollerup ◽  
Christine Elmeskov ◽  
Heidi Gumpert ◽  
Mette Pinholt ◽  
Tobias Steen Sejersen ◽  
...  
Keyword(s):  
Cell Wall ◽  
Enterococcus Faecium ◽  
Chromosomal Rearrangements ◽  
Resistance Mechanisms ◽  
Resistant Isolate ◽  
Wall Structure ◽  
Whole Genome ◽  
Cyclic Lipopeptide ◽  
Vancomycin Resistant

AbstractBackgroundDaptomycin is a cyclic lipopeptide used in the treatment of vancomycin-resistant Enterococcus faecium (VREfm). However, the development of daptomycin-resistant VREfm challenges the treatment of nosocomial VREfm infections. Resistance mechanisms of daptomycin are not fully understood. Here we analysed the genomic changes leading to a daptomycin-susceptible VREfm isolate becoming resistant after 40 days of daptomycin and linezolid combination therapy.MethodsThe two isogenic VREfm isolates (daptomycin-susceptible and daptomycin-resistant) were analysed using whole genome sequencing with Illumina and Nanopore.ResultsWhole genome comparative analysis identified the loss of a 46.5 kb fragment and duplication of a 29.7 kb fragment in the daptomycin-resistant isolate, with many implicated genes involved in cell wall synthesis. Two plasmids of the daptomycin-susceptible isolate were also found integrated in the chromosome of the resistant isolate. One nonsynonymous SNP in the rpoC gene was identified in the daptomycin-resistant isolate.ConclusionsDaptomycin resistance developed through chromosomal rearrangements leading to altered cell wall structure. Such novel types of resistance mechanisms can only be identified by comparing closed genomes of isogenic isolates.

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