Importance of the RNA secondary structure for the relative accumulation of clustered viral microRNAs
نویسندگان
چکیده
Micro (mi)RNAs are small non-coding RNAs with key regulatory functions. Recent advances in the field allowed researchers to identify their targets. However, much less is known regarding the regulation of miRNAs themselves. The accumulation of these tiny regulators can be modulated at various levels during their biogenesis from the transcription of the primary transcript (pri-miRNA) to the stability of the mature miRNA. Here, we studied the importance of the pri-miRNA secondary structure for the regulation of mature miRNA accumulation. To this end, we used the Kaposi's sarcoma herpesvirus, which encodes a cluster of 12 pre-miRNAs. Using small RNA profiling and quantitative northern blot analysis, we measured the absolute amount of each mature miRNAs in different cellular context. We found that the difference in expression between the least and most expressed viral miRNAs could be as high as 60-fold. Using high-throughput selective 2'-hydroxyl acylation analyzed by primer extension, we then determined the secondary structure of the long primary transcript. We found that highly expressed miRNAs derived from optimally structured regions within the pri-miRNA. Finally, we confirmed the importance of the local structure by swapping stem-loops or by targeted mutagenesis of selected miRNAs, which resulted in a perturbed accumulation of the mature miRNA.
منابع مشابه
Effect of Temperature on Symptoms Expression and Viral RNA Accumulation in Groundnut Bud Necrosis Virus Infected Vigna unguiculata
Background: Groundnut bud necrosis virus (GBNV) (Tospovirus genus, Bunyaviridae family) infects the major crops of solanaceae, leguminosae and cucurbitaceae in India. Temperature is an important factor which influences the plant growth and development under diseased conditions.Objective: In the present study, we evaluated the effect of four different temp...
متن کاملCOVID-19; History, Taxonomy, and Diagnostic Molecular and Immunological Techniques
In recent years, viral epidemics such as the coronavirus disease (COVID-19) have spread, and this outbreak is thought to be the result of animal-to-human transmission. Hence, accurate diagnostic tests to detect COVID-19 and antiviral antibodies in infected individuals are of utmost importance. This report describes the structure, history, taxonomy, and molecular and immunological techniques for...
متن کاملRelation Between RNA Sequences, Structures, and Shapes via Variation Networks
Background: RNA plays key role in many aspects of biological processes and its tertiary structure is critical for its biological function. RNA secondary structure represents various significant portions of RNA tertiary structure. Since the biological function of RNA is concluded indirectly from its primary structure, it would be important to analyze the relations between the RNA sequences and t...
متن کاملRNA secondary structure and qRT-PCR analyses pertained to expressed anti-CD25 CAR in NK-92 cell line
Background and Objectives: Tumor-infiltrating regulatory T (TI-Treg) cells perform the significant function in cancer immune escape. In this study, the third generation CAR construct was designed against human CD25 antigen, the significant cell surface biomarker of TI-Tregs. Methods: Initially, the construct of anti-CD25 CAR was designed. Using RNAfold web server, the RNA secondary structure wa...
متن کاملTherapeutic approaches in patients with β-thalassemia
Beta-thalassemia (β-thal) is a congenital hemoglobinopathy explained by a decreased level (β+) or absence (βο) of β-globin gene expression. Microcytic hypochromic anemia and various clinical symptoms comprising severe anemia to clinically nonsymptomatic features. Treatment with an ordered blood transfusion and iron chelator agents can decrease transfusion iron overload that causes normal matura...
متن کامل