ИСТИНА

Цель исследования – качественно и количественно охарактеризовать изменения, происходящие в транслятоме (совокупности транслируемых участков мРНК) штаммов дрожжей, нокаутных или мутантных по ряду генов, ассоциированных с процессом биосинтеза белка и скоростью репликативного старения; в случае генов с неизвестной функцией - по дифференциальной экспрессии генов и изменению метагенного профиля выдвинуть предположения о возможных функциях; в случае генов, влияющих на репликативное старение, сравнить эти изменения с теми, которые выявляются при сравнении транслятомов млекопитающих, подвергнутых продлевающим жизнь интервенциям.

The explosive growth of a field of next generation sequencing provided opportunity for a rapid comparison of the pathological phenotypes with abnormalities in specific genetic loci. However, the sharp increase in the volume of molecular genetic data, which has occurred in the last decade, is far ahead of the functional characterization of the products of the genes, whose mutations are associated with the studied pathologies. This is true not only for diseases in their classical meaning (for example, those caused by disorders in the development and functioning of the brain, including neurodegenerative diseases and autism spectrum disorders, which are now coming to the forefront in developed countries), but also more fine things - in particular, individual differences in life expectancy, rate of biological aging and age of acquisition of the first chronic diseases (healthspan). Curiously, a number of genes associated simultaneously with the aforementioned pathologies and with the difference in lifespan of mammals, encode products related to protein biosynthesis, ribosomes, or translation regulation (including genes of still unknown or poorly characterized functions). Apparently, this coincidence has a solid basis: it points to the importance of fine regulation of protein biosynthesis for maintaining cellular homeostasis, even minor disturbances of which strongly affect the complex processes of brain activity, and, in the long run, affect the rate of damage accumulation in cells, accelerating aging. Our group has been studying various translational factors for several years. Recently, we were able to discover the function of three eukaryotic proteins by successfully using the yeast model, ribosomal profiling, and classical molecular biological approaches (Young, Makeeva et al., Mol Cell 2018). Interestingly, mutations in the genes of two of these proteins (DENR and eIF2D) are associated with human brain pathologies (schizophrenia, autism, Asperger's syndrome, etc.), while the third (MCT-1) is associated with cancer. Another area of interest and competencies of our laboratory is the study of the relationship of protein biosynthesis with aging: it turns out that inhibition of the functions of various components of the translational apparatus, including that obtained by genetic manipulations, prolongs the life of many model organisms (see Anisimova et al., Aging 2018). In this project, we are going to apply the strategy, which was developed and successfully used by us in the above-mentioned study, to a number of new interesting targets related to aging and human neurodegenerative diseases, which at the same time represent components of the translation apparatus. Since the functions of these components are usually conservative within eukaryotes, we will conduct our research on yeast models. Using a systems approach (ribosomal profiling and transcriptomics), we are going to characterize functional disorders that occur during protein biosynthesis in yeast strains carrying deletions and mutations of a number of genes that are similar to those found in genetically caused diseases of the central nervous system of a person, including neurodegenerative ones and age-associated, as well as in strains with a deficiency of translational components, leading to a decrease in yeast replicative lifespan (it is well known that many genes changing this parameter in yeast, also affect the rate of aging in higher organisms). We have all these strains in our collection. The specificity of our methods (including the preparation of molecular libraries and next-generation sequencing) makes it easy to transfer them to a research platform based on the University at “Sirius” and adapt them for undergraduate and graduate students.

Будут получены и проанализированы данные рибосомного профайлинга как минимум 8 дрожжевых штаммов (wt и 7 мутантных).

С.н.с. НИИ ФХБ МГУ, доцент ФББ МГУ, к.б.н. С.Е.Дмитриев вот уже на протяжении 20 лет успешно занимается изучением биосинтеза белка в клетках эукариот. На эту тему им опубликовано более 50 статей в научных журналах, включая такие, как Nat Struct Mol Biol, PNAS, Mol Cell, Cell Rep, NAR и др. С.Е.Дмитриев является автором для корреспонденции в недавней работе в Mol Cell, описывающей применение подхода, аналогичного описанному в данном проекте, для выяснения функции трёх трансляционных факторов, eIF2D/TMA64, DENR/TMA22 и MCT-1/TMA20 (см. также ниже). Под его руководством были также выполнены другие работы по сходной тематике. Последние три года С.Е.Дмитриев является со-руководителем лаборатории системной биологии старения, созданной на средства "мегагранта" (срок которого истекает в 2019 году) совместно с проф. Гарвардской школы медицины, проф. ФББ МГУ В.Н.Гладышевым. В сотрудничестве с В.Н.Гладышевым и другими членами лаборатории им были выполнены работы по анализу изменений биосинтеза белка при старении (основная статья с описанием этих данных сейчас находится на стадии рецензирования в высокорейтинговом журнале). Это сотрудничество позволило С.Е.Дмитриеву сильно расширить свой кругозор, распространив сферу интересов на область изучения старения и контроля продолжительности жизни, что отражено в недавних публикациях двух обзоров (Aging 2018 b Ageing Res Rev 2019).