MicroRNAs (miRs) are small non-pro tein-en cod ing RNAs of 20-30 nu cleo tides long. In eukaryotic cells miRs play the role of bioregulators of gene ex pres sion through the mech a nisms of trans la tion re pression/mod u la tion. Here we both fa mil iar ize the read ers with miRs biogenesis, func tion ing mech a nisms, and strat egy of their dis cov ery and pres ent the list of bi o log i cal pro cesses, reg u lated by miRs. We also list the pub li ca tions, ded i cated to miRs role in hu man pa thol o gies (carcinogenesis in par tic u lar) and their ap pli cation for mark ing, pre ven tion, di ag nos tics, and ther apy of can cer.
The In sti tute of Mo lec u lar Bi ol ogy and Ge net ics 150 Zabolotny Str., Kyiv 03143, Ukraine The immunobiotechnological sci en tific in dus trial en ter prise, Joint-stock com pany «Diaproph-Med»
Aim. B. mori nuclear polyhedrosis virus (NPV) codes two very late genes – polyhedrin (ph) and p10. Search for miRs genes in these regions is of interest because the polyhedra, formed at the very late stage of the virus development, include small RNA of 50–60 nt. The present work was aimed at search for potential precursors of miR transcribed from the late promoter element RTAAG and the TATA promoter elements located in the ph and p10 genes regions. Methods. The search was performed using the bioinformatic programs for miR prediction: MiPred, miRNA SVM, Micropoces- sor SVM, and RNAfold. Results. It has been predicted that the region of ph gene encodes two predicted miRs (bmoNPV-miR-1ph, bmoNPV-miR-2ph) and one predicted potential (C) precursor bmoNPV- pre-miR-1Cph, which is not a Dicer substrate. The region containing p10 gene encodes one predicted miR – bmoNPV-miR- 3p10. Conclusions. A possibility of regulation of the genes orf 1629 and p74 expression by the predicted miRs, located in the same regions of a complementary chain, is assumed
It is known that B. mori NPV polyhedra contains protease which cleaves polyhedrin polypeptide chain (m.w. 28500) during dissolution of polyhedra in vitro under alkaline conditions (pH 10.5) to overlapping fragments with mol. weights 25000, 22000, 19000, 10000, and 8000. Besides, polyhedrin is present in solution at pH 10.5 as high mol. weight 13S-(70%) and >13S (30%) associates. Current paper shows that during Sepharose 6B gel-filtration at pH 10.5 of polyhedrin pH 10.5 solution there occurs partial dissociation of high mol. weight associates to unknown earlier 8.5S-associates, 25000 polypeptide and its dimer with sedimentation coefficient 3.4S. Undissociated high mol. weight associates and 8.5S-associates contain the whole polypeptide chain (m.w. 28500) only. During gel-filtration at pH 11.0 of polyhedrin solution with the same pH there occurs complete dissociation of high mol. weight associates to 8.5S-associates, 22000 polypeptide and its dimer with sedimentation coefficient 3.2S. The 8.5S-associate contains the whole 28500 polypeptide chain only. RNA was found in the low mol. weight (and high mol. weight (13S and higher) zones). 8.5S-associates do not contain RNA. After preliminary polyhedra heating at 80-90°C the polyhedra protease does not cleave polyhedrin in pH 10.5 solution and dissociation of high mol. weight associates to 8.5S-associates containing whole polypeptide chain occurs with much lower degree during gel-filtration at pH 10.5. Yet as a result of gel-filtration RNP-complex is released with mol. weight of 20000-25000 and sedimentation 2.9S, containing polypeptide chain with mol. weight of 10000-12000. The effect of the polyhedra protease and RNA on in vivo polyhedrin dissociation and virus realization from polyhedra is discussed.
В процессе растворения полиэдров вируса ядерного полиэдроза (ВЯЛ) В. mori при рН 11,0 и комнатной температуре за 2 ч протеаза полиэдров расщепляет в полипептидной цепи полиэдрина (244 остатка аминокислот) пептидные связи Tyr-Ile (208-209), Tyr-Thr (186-187), Ala-Asn (121-122) и Trp-Ser (88-89). Ни одна связь не расщепляется окончательно (100 %), по данным электрофореза в полиакриламдином геле (ПААГ) с добавлением додецилсульфата натрия, что приводит к появлению набора перекрывающихся полипептидных цепей с м.
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