Interleukin-1α (IL-1α) is a proinflammatory cytokine and a key player in host immune responses in higher eukaryotes. IL-1α has pleiotropic effects on a wide range of cell types, and it has been extensively studied for its ability to contribute to various autoimmune and inflammation-linked disorders, including rheumatoid arthritis, Alzheimer's disease, systemic sclerosis and cardiovascular disorders. Interestingly, a significant proportion of IL-1α is translocated to the cell nucleus, in which it interacts with histone acetyltransferase complexes. Despite the importance of IL-1α, little is known regarding its binding targets and functions in the nucleus. We took advantage of the histone acetyltransferase (HAT) complexes being evolutionarily conserved from yeast to humans and the yeast SAGA complex serving as an epitome of the eukaryotic HAT complexes. Using gene knock-out technique and co-immunoprecipitation of the IL-1α precursor with TAP-tagged subunits of the yeast HAT complexes, we mapped the IL-1α-binding site to the HAT/Core module of the SAGA complex. We also predicted the 3-D structure of the IL-1α N-terminal domain, and by employing structure similarity searches, we found a similar structure in the C-terminal regulatory region of the catalytic subunit of the AMP-activated/Snf1 protein kinases, which interact with HAT complexes both in mammals and yeast, respectively. This finding is further supported with the ability of the IL-1α precursor to partially rescue growth defects of snf1Δ yeast strains on media containing 3-Amino-1,2,4-triazole (3-AT), a competitive inhibitor of His3. Finally, the careful evaluation of our data together with other published data in the field allows us to hypothesize a new function for the ADA complex in SAGA complex assembly.

• MeSH
• biologické modely MeSH
• buněčné jádro metabolismus   MeSH
• genový knockout MeSH
• histonacetyltransferasy metabolismus   MeSH
• imunoprecipitace MeSH
• interleukin-1alfa chemie   metabolismus   MeSH
• lidé MeSH
• podjednotky proteinů metabolismus   MeSH
• protein-serin-threoninkinasy chemie   metabolismus   MeSH
• proteinkinasy aktivované AMP chemie   metabolismus   MeSH
• proteinové prekurzory chemie   metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny metabolismus   MeSH
• Saccharomyces cerevisiae metabolismus   MeSH
• signální transdukce MeSH
• strukturní homologie proteinů MeSH
• subcelulární frakce metabolismus   MeSH
• terciární struktura proteinů MeSH
• trans-aktivátory metabolismus   MeSH
• vazba proteinů MeSH
• vazebná místa MeSH
• výpočetní biologie MeSH
• vztahy mezi strukturou a aktivitou MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• histonacetyltransferasy MeSH
• interleukin-1alfa MeSH
• podjednotky proteinů MeSH
• protein-serin-threoninkinasy MeSH
• proteinkinasy aktivované AMP MeSH
• proteinové prekurzory MeSH
• Saccharomyces cerevisiae - proteiny MeSH
• SAGA complex, S cerevisiae MeSH Prohlížeč
• SNF1-related protein kinases MeSH Prohlížeč
• trans-aktivátory MeSH

BACKGROUND: The knowledge of biological characteristics of minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) remains sparse. There are no data available on what level of MRD might be 'safe' without an overt risk of relapse, or whether any such level exists at all. To address this issue in prospective studies, we have developed a quantitative molecular approach to monitor MRD in CLL, which allows the malignant clone to be traced with far higher sensitivity than possible with the techniques available currently. METHOD: To quantify MRD in CLL patients, a novel locked nucleic acid (LNA)-RNA-based quantitative real-time PCR technique was developed. Clone-specific assays were prepared for 62 CLL patients. Thirty patients were followed up molecularly for a median of 250 days (range 69-570 days). All patients were administered chemo/immunotherapy. RESULTS: In three patients, molecular negativity was achieved, as estimated by LNA-based assays. In one patient, a sustained molecular negativity was established by chemo/immunotherapy and the patient remains molecularly negative (322 days). The LNA-based assay enabled us to evaluate MRD in a reproducible manner with the sensitivity of 10(-7). CONCLUSION: LNA-RNA-based quantitative real-time PCR is an effective approach for MRD monitoring with the potential for increased sensitivity compared with standard DNA-based assays used for molecular follow-up.

• MeSH
• antigeny CD38 genetika   metabolismus   MeSH
• chronická lymfatická leukemie diagnóza   genetika   MeSH
• diagnostické techniky molekulární metody   MeSH
• hybridizace nukleových kyselin metody   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorový supresorový protein p53 genetika   metabolismus   MeSH
• polymerázová řetězová reakce metody   MeSH
• prospektivní studie MeSH
• protein-tyrosinkináza ZAP-70 genetika   metabolismus   MeSH
• průtoková cytometrie MeSH
• reprodukovatelnost výsledků MeSH
• reziduální nádor diagnóza   genetika   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• těžké řetězce imunoglobulinů genetika   metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD38 MeSH
• nádorový supresorový protein p53 MeSH
• protein-tyrosinkináza ZAP-70 MeSH
• těžké řetězce imunoglobulinů MeSH