Transcription factors (TFs) play a central role in the gene regulation associated with a plant's development and its response to the environmental factors. The work of TFs is well regulated at each stage of their activities. TFs usually consist of three protein domains required for DNA binding, dimerization, and transcriptional regulation. Alternative splicing (AS) produces multiple proteins with varying composition of domains. Recent studies have shown that AS of some TF genes form small proteins (small interfering peptide/small interfering protein, siPEP/siPRoT), which lack one or more domains and negatively regulate target TFs by the mechanism of protein interference (peptide interference/protein interference, PEPi/PROTi). The presence of an alternative form for the transcription factor CCA1 of Arabidopsis thaliana, has been shown to be involved in the regulation of the response to cold stress. For the PtFLC protein, one of the isoforms was found, which is formed as a result of alternative splicing and acts as a negative repressor, binding to the full-length TF PtFLC and therefore regulating the development of the Poncirus trifoliata. For A. thaliana, a FLM gene was found forming the FLM-б isoform, which acts as a dominant negative regulator and stimulates the development of the flower formation process due to the formation of a heterodimer with SVP TF. Small interfering peptides and proteins can actively participate in the regulation of gene expression, for example, in situations of stress or at different stages of plant development. Moreover, small interfering peptides and proteins can be used as a tool for fundamental research on the function of genes as well as for applied research for permanent or temporary knockout of genes. In this review, we have demonstrated recent studies related to siPEP/siPROT and their involvement in the response to various stresses, as well as possible ways to obtain small proteins.

Here we present the method for the target-specific elimination of certain intracellular proteins in plants using the ubiquitin-proteasome system. We modified the E3 ubiquitin ligase Chip of A. thaliana to obtain two variants carrying the deletions at the N-terminus and the GFP recognition domain. The interaction of the GFP protein and the chimeric ubiquitin ligase was confirmed via yeast two-hybrid assay. Fluorescence microscopy and fluorimetry showed that, when an infiltration of the gfp-expressing N. benthamiana plants was performed with agrobacteria carrying the hybrid E3 gene of the ubiquitin ligase Chip with the GFP recognition domain, a significant decrease in the fluorescence was observed for both variants: carrying the N-terminal deletion of 100 amino acids or the deletion of 140 amino acids.

Matrix metalloproteinases are crucial for the maintenance of skin homeostasis, wound healing, and fueling the inflammatory process. In psoriasis, matrix metalloproteinases contribute to epidermal remodeling. They also influence the composition of extracellular matrix and intercellular interactions. Moreover, matrix metalloproteinases are important for vasodilation of dermal microcapillaries. The aim of this paper is to clarify whether knocking matrix metalloproteinase 1 down in epidermal keratinocytes can be beneficial for psoriasis. Lentiviral transduction was used to obtain the cell lines HaCaT-IK and HaCaT-KTR that expressed shRNA specific to matrix metalloproteinase 1 and scrambled shRNA, respectively. Changes in gene expression were analyzed by qPCR. The enzyme activity of matrix metalloproteinase 1 was assessed by zymography. Scratch assay was used to assess migration in transduced cells. Comparative analysis of HaCaT-IK and HaCaT-KTR revealed that MMP1 silencing downregulated matrix metalloproteinase 1 and decreased the enzyme activity of the named enzyme in 7.5 and 4 times, respectively. However, MMP1 silencing changed neither the expression of homologous genes (MMP2, -9 and -12) nor proliferation rate of the transduced cells. Surprisingly, MMP1-silencing changed the expression of several genes that are essential for pathogenesis of psoriasis. Particularly, MMP1-silencing caused downregulation of CCNA2 (0.54 ± 0.14) and KRT17 (0.58 ± 0.18). It also led to induction of LOR (2.08 ± 0.16), FLG (3.29 ± 0.35), KRT1 -5 and -10 (2.06 ± 0.20, 1.50 ± 0.22 and 2.09 ± 0.10, respectively). Finally, MMP1-silencing suppressed cell migration on collagen. In conclusion, MMP1 silencing in human epidermal keratinocytes causes changes in expression of LOR, FLG KRT1, -5, -10 and -17 that can be beneficial for psoriasis.

FRA1, a transcription factor from AP-1 superfamily, is an important regulator of cell fate. The expression patterns of FRA1 in skin vary depending on the epidermal layer and the differentiation state of keratinocytes, and our previous research had shown FRA1 to be overexpressed in all epidermal layers of lesional skin of patients with psoriasis. The hallmarks of the disease are altered immune profiles of skin, keratinocyte hyperproliferation and abnormal differentiation, as well as extensive remodeling of extracellular matrix and keratinocyte's acquisition of mesenchymal transition phenotype. Keratinocytes seem to be important mediators of the disease, not only responsible for visible skin manifestations and structural changes, but also capable for production of vast majority of psoriasis-associated cytokines, chemokines and antimicrobial molecules. In order to evaluate the role of FRA1 regulation in psoriasis-associated phenotype of keratinocytes we have created HaCaT keratinocytes with inducible FRA1 overexpression. qPCR analysis of FRA1-overexpressing cells has shown increased expression of proteases MMP1, MMP2 and MMP12 (ECM remodeling), cytokines TNFa and IL-8 (inflammation), SLUG, FN1, SERPINE1 (mesenchymal markers), CK16 (proliferation) and decreased expression of CK10 (differentiation). The influence of FRA1 overexpression on cell migration and wound healing was evaluated by scratch assay that has shown enhanced capability of FRA1-overexpressing cells to migrate toward the scratch. Our results support the hypothesis that FRA1 is an important player in the developing of psoriasis-associated skin manifestations, contributing to the elevated cytokine production by keratinocytes. Besides, FRA1-associated MMP-mediated ECM remodeling itself could promote the acquisition of mesenchymal phenotype by keratinocytes leading to the shift of CK expression, activation of proliferation and migration of the cells in the course of the disease.

Experimental data obtained in this study had shown the involvement of A. thaliana immunophilin genes At2g16600, At4g33060, and At5g48570 in plant defense responses to the Xanthomonas campestris invasion. We had found not only that the expression levels of these genes changed upon bacterial infection, but also that the plant’s resistance to the pathogen was increased if the expression levels of the immunophilin genes were elevated in the host cells.

Proinflammatory cytokines TNF, IFNG, and IL17 play an important role in eruption of psoriasis. The activation of epidermal keratinocytes with the named cytokines alters their terminal differentiation program and causes their hyperproliferation in the diseased skin. HaCaT cells, which are immortalized human keratinocytes, are often used as a cellular model of psoriasis. The aim of this study was to evaluate changes in gene expression and the proliferation rates in cultured HaCaT cells treated with TNF, IFNG, and IL17. We found that HaCaT cells decrease their proliferation rate in response to either IL17 or a combination TNF and IFNG. The analysis of microarray data discovered a group of 12 genes, which were downregulated in HaCaT after treatments with the named cytokines and upregulated in psoriatic lesional skin. Eight genes were important for DNA replication and they also contributed to two larger networks that regulated cell progression through the cell cycle. We conclude that HaCaT cells have a sufficient limitation as a cellular model of psoriasis due to their treatment with proinflammatory cytokines, namely TNF, IFNG, and IL17 does not increase their proliferation rate. Thus, the studies of psoriasis based on HaCaT cells as an experimental model shall take in account this important phenomenon.

DOI of original article: http://dx.doi.org/10.1016/j.gene.2014.01.029. ⁎ Corresponding author at: 219 Bessey Hall, Department of Plant Pathology and Microbiology, Iowa State University, Ames 50014, IA, USA. Tel.: +1 515 294 3120. E-mail addresses: gennady@iastate.edu, gpogorelko@yandex.ru (G.V. Pogorelko), marja-2007@yandex.ru (M. Mokryakova), oksfursova@yandex.ru (O.V. Fursova), insaz@yandex.ru (I. Abdeeva), eleopiru@vigg.ru (E.S. Piruzian), sergey.bruskin@gmail.com (S.A. Bruskin).

Plant immunophilins are a broadly conserved family of proteins, which carry out a variety of cellular functions. In this study, we investigated three immunophilin genes involved in the Arabidopsis thaliana response to Pseudomonas syringae infection: a cytoplasmic localized AtCYP19, a cytoplasmic and nuclear localized AtCYP57, and one nucleus directed FKBP known as AtFKBP65. Arabidopsis knock-out mutations in these immunophilins result in an increased susceptibility to P. syringae, whereas overexpression of these genes alters the transcription profile of pathogen-related defense genes and led to enhanced resistance. Histochemical analysis revealed local gene expression of AtCYP19, AtCYP57, and AtFKBP65 in response to pathogen infection. AtCYP19 was shown to be involved in reactive oxygen species production, and both AtCYP57 and AtFKBP65 provided callose accumulation in plant cell wall. Identification of the involvement of these genes in biotic stress response brings a new set of data that will advance plant immune system research and can be widely used for further investigation in this area.

Провоспалительные цитокины: фактор некроза опухолей (ФНО), интерферон (ИФНГ) и интерлейкин 17 (ИЛ-17) – играют важную роль в патогенезе псориаза. Активация эпидермальных кератиноцитов этими цитокинами изменяет процесс дифференцировки клеток, что является основной причиной их гиперпролиферации в коже больных. Иммортализованную линию кератиноцитов человека HaCaT часто используют в качестве модельной системы для изучения молекулярных механизмов патогенеза псориаза. Установлено, что обработка клеток HaCaT провоспалительными цитокинами приводит к снижению пролиферации этих клеток. Анализ генной экспрессии выявил группу из 12 генов, экспрессия которых повышена при псориазе, но снижена в клетках HaCaT после их обработки цитокинами. Выявленные гены играют важную роль в процессе репликации ДНК и входят в состав двух других, более обширных групп генов с измененной экспрессией, участвующих в регуляции клеточного цикла. Полученные в ходе данной работы результаты свидетельствуют о том, что культура клеток HaCaT имеет существенное ограничение в качестве модельной системы псориаза: обработка HaCaT провоспалительными цитокинами не приводит к увеличению пролиферации клеток. Эту особенность необходимо принимать во внимание при изучении влияния новых лекарственных препаратов на пролиферацию клеток.

Receptor for advanced glycation end-products is implicated in a development of chronic inflammatory response. Aim of this paper is to provide a review on commercial and experimental medicines that can interfere with RAGE and signaling through RAGE. We searched three bibliographical databases (PubMed, Web of Science and MEDLINE) for the publications from 2005 to March 2012 and identified 5 major groups of agents that can interfere with RAGE biological effects. In the first part of this paper, we discuss AGE crosslink breakers. These chemicals destroy advanced glycation end products (AGEs) that are crosslinked to the extracellular matrix proteins and can interact with RAGE as ligands. Then, we describe two non-conventional agents SAGEs and KIOM-79 that abolish certain biological effects of RAGE and have a strong anti-inflammatory potential. In the third part, we evaluate the inhibitors of the signaling cascades that underlie RAGE. Particularly, we discuss two groups of kinase inhibitors tyrphostins and the inhibitors of JAK kinases. Considering RAGE as a potential master regulator of processes that are crucial for the pathogenesis of psoriasis, we propose that these medicins may help in controlling the disease by abolishing the chronic inflammation in skin lesions.

Одним из главных инструментов патогенеза грамотрицательных бактерий являются белки, секретируемые при помощи транспортной системы третьего типа (T3TS) внутрь растительной клетки. Класс белков, доставляемых в растительную клетку через T3TS, имеет общее название белков-эффекторов. На основе анализа геномных последовательностей восьми полностью секвенированных геномов ксантомонад нами был определен состав потенциальных генов белков-эффекторов, куда вошли 19 генов как с известными, так и с неизвестными функциями. Методом ПЦР был определен состав генов белков-эффекторов, характерный для каждого штамма ксантомонад в исследуемой популяции. Был показан высокий уровень генетической изменчивости состава генов-эффекторов в пределах одного вида и его патовариантов. Впервые, у ряда штаммов Xanthomonas campestris pv. campestris были обнаружены гены-эффекторы, несвойственные типовым штаммам этого вида, и выдвинуто предположение о связи ряда генов с симптомами вызываемых поражений на растении-хозяине, а также показана связь числа генов-эффекторов с основными расами патогена.

Psoriasis was used as a model to analyze the pathogenetic pathways of immune-mediated inflammatory diseases, and the results of bioinformatic, molecular-genetic and proteomic studies are provided. Cell mechanisms, common for the pathogenesis of psoriasis, as well as Crohn’s disease, are identified. New approaches for immune-mediated diseases are discussed.

Проведен сравнительный биоинформатический анализ молекулярно-генетических процессов в патогенезе многофакторных заболеваний – псориаза и болезни Крона – на основании обработки данных биологических микрочипов базы данных GEO DataSets. Установлены общие гены и общие молекулярно-генетические процессы для двух патологий. Сделано предположение об участии ряда транскрипционных факторов, включая систему транскрипционных факторов АР-1, в патогенезе как псориаза, так и болезни Крона.

A comparative bioinformatics analysis of the molecular genetic processes in the pathogenesis of multifactorial diseases-psoriasis and Crohn's disease-was performed using the results of microarray experiments deposited with the GEO DataSets database. A number of genes and molecular genetic processes common to both pathologies were found. The involvement of several transcription factors, including AP-1 transcription factors, in the pathogeneses of both psoriasis and Crohn's disease was postulated.