Cellular senescence guards against cancer and modulates aging; however, the underlying mechanisms remain poorly understood. Here, we show that genotoxic drugs capable of inducing premature senescence in normal and cancer cells, such as 5-bromo-2'-deoxyuridine (BrdU), distamycin A (DMA), aphidicolin and hydroxyurea, persistently activate Janus kinase-signal transducer and activator of transcription (JAK/STAT) signaling and expression of interferon-stimulated genes (ISGs), such as MX1, OAS, ISG15, STAT1, PML, IRF1 and IRF7, in several human cancer cell lines. JAK1/STAT-activating ligands, interleukin 10 (IL10), IL20, IL24, interferon gamma (IFNgamma), IFNbeta and IL6, were also expressed by senescent cells, supporting autocrine/paracrine activation of JAK1/STAT. Furthermore, cytokine genes, including proinflammatory IL1, tumor necrosis factor and transforming growth factor families, were highly expressed. The strongest inducer of JAK/STAT signaling, cytokine production and senescence was BrdU combined with DMA. RNA interference-mediated knockdown of JAK1 abolished expression of ISGs, but not DNA damage signaling or senescence. Thus, although DNA damage signaling, p53 and RB activation, and the cytokine/chemokine secretory phenotype are apparently shared by all types of senescence, our data reveal so far unprecedented activation of the IFNbeta-STAT1-ISGs axis, and indicate a less prominent causative role of IL6-JAK/STAT signaling in genotoxic drug-induced senescence compared with reports on oncogene-induced or replicative senescence. These results highlight shared and unique features of drug-induced cellular senescence, and implicate induction of cancer secretory phenotype in chemotherapy.
- MeSH
- bromodeoxyuridin farmakologie MeSH
- cytokiny genetika metabolismus MeSH
- distamyciny farmakologie MeSH
- HeLa buňky MeSH
- interferony genetika metabolismus MeSH
- interleukin-10 genetika metabolismus MeSH
- interleukin-6 genetika metabolismus MeSH
- interleukin-8 genetika metabolismus MeSH
- interleukiny genetika metabolismus MeSH
- Janus kinasa 1 genetika metabolismus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- RNA interference MeSH
- signální transdukce účinky léků MeSH
- stárnutí buněk účinky léků MeSH
- synergismus léků MeSH
- transkripční faktor STAT1 genetika metabolismus MeSH
- western blotting MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bromodeoxyuridin MeSH
- cytokiny MeSH
- distamyciny MeSH
- interferony MeSH
- interleukin 20 MeSH Prohlížeč
- interleukin-10 MeSH
- interleukin-24 MeSH Prohlížeč
- interleukin-6 MeSH
- interleukin-8 MeSH
- interleukiny MeSH
- Janus kinasa 1 MeSH
- stallimycin MeSH Prohlížeč
- STAT1 protein, human MeSH Prohlížeč
- transkripční faktor STAT1 MeSH
The requirement for novel platinum antitumor drugs led to the concept of synthesis of novel platinum drugs based on targeting cisplatin to various carrier molecules. We have shown [Loskotova, H., and Brabec, V. (1999) Eur. J. Biochem. 266, 392-402] that attachment of DNA minor-groove-binder distamycin to cisplatin changes several features of DNA-binding mode of the parent platinum drug. Major differences comprise different conformational changes in DNA and a considerably higher interstrand cross-linking efficiency. The studies of the present work have been directed to the analysis of oligodeoxyribonucleotide duplexes containing single, site-specific adducts of platinum-distamycin conjugates. These uniquely modified duplexes were analyzed by Maxam-Gilbert footprinting, phase-sensitive gel electrophoresis bending assay and chemical probes of DNA conformation. The results have indicated that the attachment of distamycin to cisplatin mainly affects the sites involved in the interstrand cross-links so that these adducts are preferentially formed between complementary guanine and cytosine residues. This interstrand cross-link bends the helix axis by approximately 35 degrees toward minor groove, unwinds DNA by approximately 95 degrees and distorts DNA symmetrically around the adduct. In addition, CD spectra of restriction fragments modified by the cisplatin-distamycin conjugates have demonstrated that distamycin moiety in the interstrand cross-links of these compounds interacts with DNA. This interaction facilitates the formation of these adducts. Hence, the structural impact of the specific interstrand cross-link detected in this study deserves attention when biological behavior of cisplatin derivatives targeted by oligopeptide DNA minor-groove-binders is evaluated.
- MeSH
- adukty DNA chemie MeSH
- brom chemie MeSH
- cirkulární dichroismus MeSH
- cisplatina chemie MeSH
- diethylpyrokarbonát chemie MeSH
- dinukleosidfosfáty chemie MeSH
- distamyciny chemie MeSH
- DNA chemie MeSH
- heteroduplexy nukleové kyseliny chemická syntéza chemie MeSH
- jednovláknová DNA chemie MeSH
- konformace nukleové kyseliny * MeSH
- manganistan draselný chemie MeSH
- oligodeoxyribonukleotidy chemická syntéza chemie MeSH
- pyrimidinové nukleotidy chemická syntéza chemie MeSH
- reagencia zkříženě vázaná chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adukty DNA MeSH
- brom MeSH
- cisplatin-deoxy(guanosine monophosphate guanosine) adduct MeSH Prohlížeč
- cisplatin-DNA adduct MeSH Prohlížeč
- cisplatina MeSH
- deoxyguanylyl-(3'-5')-guanosine MeSH Prohlížeč
- diethylpyrokarbonát MeSH
- dinukleosidfosfáty MeSH
- distamyciny MeSH
- DNA MeSH
- heteroduplexy nukleové kyseliny MeSH
- jednovláknová DNA MeSH
- manganistan draselný MeSH
- oligodeoxyribonukleotidy MeSH
- pyrimidinové nukleotidy MeSH
- reagencia zkříženě vázaná MeSH
- stallimycin MeSH Prohlížeč
Modifications of natural DNA in a cell-free medium using cisplatin tethered to the AT-specific, minor groove binder distamycin, were studied using various methods of biochemical analysis or molecular biophysics. These methods include: binding studies using differential pulse polarography and flameless atomic absorption spectrophotometry, mapping DNA adducts using a transcription assay, use of ethidium bromide as a fluorescent probe for DNA adducts of platinum, measurement of DNA unwinding by gel electrophoresis, measurement of CD spectra, an interstrand cross-linking assay using gel electrophoresis under denaturing conditions, measurement of melting curves with the aid of absorption spectrophotometry and the use of terbium ions as a fluorescent probe for distorted base pairs in DNA. The results indicate that attachment of distamycin to cisplatin changes several features of the DNA-binding mode of the parent platinum drug. Major differences comprise different conformational alterations in DNA and a considerably higher efficiency of the conjugated drug to form in DNA interstrand cross-links. Cisplatin tethered to distamycin, however, coordinates to DNA with similar base sequence preferences as the untargeted platinum drug. The results point to a unique profile of DNA binding for cisplatin-distamycin conjugates, suggesting that tethering cisplatin to minor groove oligopeptide binders may also lead to an altered biological activity profile.
- MeSH
- adukty DNA * MeSH
- antitumorózní látky farmakologie MeSH
- bezbuněčný systém MeSH
- časové faktory MeSH
- cirkulární dichroismus MeSH
- cisplatina farmakologie MeSH
- distamyciny farmakologie MeSH
- DNA vazebné proteiny chemie MeSH
- DNA chemie metabolismus ultrastruktura MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- ethidium farmakologie MeSH
- genetická transkripce MeSH
- interkalátory farmakologie MeSH
- konformace nukleové kyseliny MeSH
- molekulární modely MeSH
- peptidy chemie MeSH
- reagencia zkříženě vázaná farmakologie MeSH
- sekvence nukleotidů MeSH
- superhelikální DNA MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- adukty DNA * MeSH
- antitumorózní látky MeSH
- cisplatina MeSH
- distamyciny MeSH
- DNA vazebné proteiny MeSH
- DNA MeSH
- ethidium MeSH
- interkalátory MeSH
- peptidy MeSH
- reagencia zkříženě vázaná MeSH
- stallimycin MeSH Prohlížeč
- superhelikální DNA MeSH
A single-strand-specific chemical probe, potassium permanganate (KMnO4), was used to study the sequence-dependent conformation periodicity of tandem multicopy repetitive DNA sequences HRS60 and GRS (Nicotiana Species) at the level of single base pair and dinucleotide step. Local DNA structures, sensitive to KMnO4, revealed periodicity of 182 +/- 2 bp, equal to the length of repeat units. Permanganate-sensitive local structures were mapped to both DNA strands of genomic HRS60 sequences and were found to be linked to d(A)n tracts. These adenine tracts are located in the proximity of the intrinsically curved domains. Distamycin A increased reactivity of the DNA but decreased the specificity of DNA cleavage. Similar conformation periodicity has been detected also in the 'canrep' family of repeats (Brassica species). All studied repetitive sequences are predominantly located in the constitutive heterochromatin. We discuss the role of conformation periodicities in relation to a structural code for nucleosome phasing at tandem arrays of DNA repeats.
- MeSH
- Brassica genetika MeSH
- distamyciny MeSH
- DNA primery genetika MeSH
- DNA rostlinná chemie genetika MeSH
- jedovaté rostliny MeSH
- konformace nukleové kyseliny * MeSH
- manganistan draselný MeSH
- mapování chromozomů MeSH
- molekulární sekvence - údaje MeSH
- molekulární sondy MeSH
- polymorfismus genetický MeSH
- repetitivní sekvence nukleových kyselin * MeSH
- sekvence nukleotidů MeSH
- tabák genetika MeSH
- vazebná místa MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- distamyciny MeSH
- DNA primery MeSH
- DNA rostlinná MeSH
- manganistan draselný MeSH
- molekulární sondy MeSH
- stallimycin MeSH Prohlížeč
Repetitive basic polypeptides containing lysine or arginine as every third amino acid were shown to cause DNA condensation at physiological salt concentration connected with selective DNA binding with respect to DNA composition and sequence. This selectivity is very similar to that existing in the case of histone H1 and other basic proteins and does not depend on polypeptide chain conformation. The effect of the minor groove binding drugs netropsin and distamycin was tested to elucidate the origin of the binding selectivity. The results suggest that the binding preferences are due to the variations in the conformation in various types of B-DNA that depend on DNA composition and sequence. The most important factor affecting the selectivity is probably the value of the negative electrostatic potential in the minor groove.
- MeSH
- chemické modely MeSH
- distamyciny chemie metabolismus MeSH
- DNA chemie metabolismus MeSH
- konformace nukleové kyseliny MeSH
- konformace proteinů MeSH
- netropsin chemie metabolismus MeSH
- peptidy chemická syntéza chemie metabolismus MeSH
- vazba proteinů * MeSH
- vazebná místa MeSH
- zastoupení bazí MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- distamyciny MeSH
- DNA MeSH
- netropsin MeSH
- peptidy MeSH
- stallimycin MeSH Prohlížeč
Structural properties and length distribution profile of 7 S avian myeloblastosis virus (AMV) DNA were studied by means of electron microscopy using two different techniques. This DNA represents mostly double strands, the single strands being in minority. We have shown directly that this DNA forms a bent structure typical of the majority of molecules. These bends are sensitive to the distamycin treatment which stretches most of the bent molecules. Some amount (up to 30%) of circular DNA molecules was detected also in DNA preparations, the nature and the size of which are reminiscent of electron microscopic data on microbubbles of replicating DNA. No specific AMV DNA structural features were found using osmium-tetroxide treatment. The basic size of AMV DNA was estimated to be approximately 150 bp, but its multimers were also detected. Their presence and significance is discussed.
- MeSH
- distamyciny farmakologie MeSH
- DNA virů chemie účinky léků ultrastruktura MeSH
- elektronová mikroskopie MeSH
- jednovláknová DNA chemie ultrastruktura MeSH
- konformace nukleové kyseliny účinky léků MeSH
- kruhová DNA chemie ultrastruktura MeSH
- kur domácí MeSH
- molekulární struktura MeSH
- ptačí leukóza mikrobiologie MeSH
- retrovirové infekce mikrobiologie MeSH
- virus ptačí myeloblastózy chemie izolace a purifikace ultrastruktura MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- distamyciny MeSH
- DNA virů MeSH
- jednovláknová DNA MeSH
- kruhová DNA MeSH
