The accessions of the morel (Morchellaceae, Ascomycota) germplasm collection were genetically analyzed, in order to determine both their inter- and intraspecific relationships. This was done as a starting point for cultivation experiments, as well as to provide a genetic description of invasive morel populations linked to mulched garden patches, as compared with outdoor morels. The phylogenetic data, which was based on the internal transcribed spacer (ITS) sequences and supported by amplified fragment length polymorphism (AFLP) analyses, divided the germplasm isolates and accessions from the sequence database into three groups of yellow morels, and three groups of black morels, involving a remarkable monotypic genus of half-free morels (Mitrophora semilibera), the groups Morchella conica and M. angusticeps. Both Morchella groups include morel samples that use mulch bark as a vector for their spread across gardens in various locations in the Czech Republic. The AFLP analysis supported the ITS-based phylogenetic data and determined the intraspecific genetic profile of these, as a rule, almost entirely unstudied isolates.
- MeSH
- analýza polymorfismu délky amplifikovaných restrikčních fragmentů MeSH
- Ascomycota klasifikace genetika izolace a purifikace MeSH
- DNA fungální genetika MeSH
- funkční potraviny analýza klasifikace MeSH
- fylogeneze MeSH
- intergenová DNA genetika MeSH
- molekulární sekvence - údaje MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
In this work, we have studied the structural and functional linkage between lamin A/C, nuclear actin, and organization of chromosome territories (CTs) in mammary carcinoma MCF-7 cells. Selective down-regulation of lamin A/C expression led to disruption of the lamin A/C perinuclear layer and disorganization of lamin-bound emerin complexes at the inner nuclear membrane. The silencing of lamin A/C expression resulted in a decrease in the volume and surface area of chromosome territories, especially in chromosomes with high heterochromatin content. Inhibition of actin polymerization led to relaxation of the structure of chromosome territories, and an increase in the volumes and surface areas of the chromosome territories of human chromosomes 1, 2 and 13. The results show an important role of polymeric actin in the organization of the nuclei and the chromosome territories.
- MeSH
- aktiny metabolismus MeSH
- buněčné jádro metabolismus MeSH
- down regulace MeSH
- financování organizované MeSH
- genom lidský genetika MeSH
- konfokální mikroskopie MeSH
- lamin typ A metabolismus MeSH
- lidé MeSH
- lidské chromozomy metabolismus MeSH
- nádorové buněčné linie MeSH
- tvar buňky MeSH
- Check Tag
- lidé MeSH
Functions of nuclear polymeric proteins such as lamin A/C and actin in transport of plasmid DNA were studied. The results show that the lamina plays an important role in plasmid DNA's entry into the cell nucleus from the cytoplasm. Selective disruption of lamin A/C led to a halt in plasmid DNA transport through the nuclear envelope. Inside the nucleus, plasmid DNA was frequently localized at sites with impaired genome integrity, such as DNA double-strand breaks (DSBs), occurring spontaneously or induced by ionizing radiation. Polymeric actin obviously participates in nuclear transport of plasmid DNA, since inhibition of actin polymerization by latrunculin B disturbed plasmid transport inside the cell nucleus. In addition, precluding of actin polymerization inhibited plasmid co-localization with newly induced DSBs. These findings indicate the crucial role of polymeric actin in intranuclear plasmid transport.
- MeSH
- aktiny genetika chemie metabolismus MeSH
- aktivní transport - buněčné jádro MeSH
- buněčné jádro metabolismus MeSH
- buněčné linie MeSH
- dvouřetězcové zlomy DNA MeSH
- financování organizované MeSH
- laminy MeSH
- lidé MeSH
- plazmidy genetika metabolismus MeSH
- rekombinantní DNA genetika metabolismus MeSH
- rekombinantní proteiny genetika chemie metabolismus MeSH
- transfekce MeSH
- umlčování genů MeSH
- záření gama MeSH
- Check Tag
- lidé MeSH
We show that double strand breaks (DSBs) induced in chromatin of low as well as high density by exposure of human cells to gamma-rays are repaired in low-density chromatin. Extensive chromatin decondensation manifested in the vicinity of DSBs by decreased intensity of chromatin labelling, increased H4K5 acetylation, and decreased H3K9 dimethylation was observed already 15 min after irradiation. Only slight movement of sporadic DSB loci for short distances was noticed in living cells associated with chromatin decondensation around DSBs. This frequently resulted in their protrusion into the low-density chromatin domains. In these regions, the clustering (contact or fusion) of DSB foci was seen in vivo, and in situ after cell fixation. The majority of these clustered foci were repaired within 240 min, but some of them persisted in the nucleus for several days after irradiation, indicating damage that is not easily repaired. We propose that the repair of DSB in clustered foci might lead to misjoining of ends and, consequently, to exchange aberrations. On the other hand, the foci that persist for several days without being repaired could lead instead to cell death.
- MeSH
- acetylace MeSH
- buněčné linie MeSH
- chromatin metabolismus MeSH
- dvouřetězcové zlomy DNA MeSH
- fibroblasty metabolismus účinky záření MeSH
- financování organizované MeSH
- histony metabolismus MeSH
- lidé MeSH
- metylace MeSH
- nádorové buněčné linie MeSH
- oprava DNA MeSH
- záření gama MeSH
- Check Tag
- lidé MeSH
This work is focused on the function of the microtubule and actin networks in plasmid DNA transport during liposomal transfection. We observed strong binding of plasmid DNA-lipid complexes (lipoplexes) to both networks and directional long-range motion of these lipoplexes along the microtubules. Disruption of either of these networks led to the cessation of plasmid transport to the nucleus, a decreased mobility of plasmids, and accumulation of plasmid DNA in large aggregates at the cell periphery. Our findings show an indispensable but different role of both types of cytoskeleton, actin and microtubular, in the processes of gene delivery.
- MeSH
- aktiny fyziologie metabolismus MeSH
- bicyklické sloučeniny heterocyklické farmakologie MeSH
- biologický transport účinky léků MeSH
- cytochalasin D farmakologie MeSH
- DNA genetika metabolismus MeSH
- fibroblasty cytologie metabolismus účinky léků MeSH
- financování organizované MeSH
- imunoblotting MeSH
- kultivované buňky MeSH
- lidé MeSH
- mikrotubuly fyziologie metabolismus MeSH
- plazmidy genetika metabolismus MeSH
- rekombinantní fúzní proteiny genetika metabolismus MeSH
- thiazolidiny farmakologie MeSH
- transfekce MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
Movement of labelled plasmid DNA relative to heterochromatin foci in nuclei, visualized with HP1-GFP, was studied using live-cell imaging and object tracking. In addition to Brownian motion of plasmid DNA we found a pronounced, non-random movement of plasmid DNA towards the nearest HP1 focus, while time-lapse microscopy showed that HP1 foci are relatively immobile and positionally stable. The movement of plasmid DNA was much faster than that of the HP1 foci. Contact of transgene DNA with an HP1 focus usually resulted in cessation of the directional motion. Moreover, the motion of plasmid DNA inside the heterochromatin compartment was more restricted (limited to 0.25 microm) than when the plasmid DNA was outside heterochromatin (R = 0.7 microm). Three days after transfection most of the foreign labelled DNA colocalized with centromeric heterochromatin.
- MeSH
- biologický transport fyziologie genetika MeSH
- buněčné jádro fyziologie MeSH
- chromozomální proteiny, nehistonové fyziologie genetika MeSH
- DNA fyziologie genetika MeSH
- financování organizované MeSH
- heterochromatin MeSH
- lidé MeSH
- mikroskopie MeSH
- nádorové buněčné linie MeSH
- plazmidy fyziologie genetika MeSH
- transfekce MeSH
- Check Tag
- lidé MeSH
- MeSH
- granulocyty MeSH
- heterochromatin MeSH
- histony MeSH
- krevní buňky MeSH
- lidé MeSH
- metylace MeSH
- myeloidní leukemie MeSH
- Check Tag
- lidé MeSH
