scholarly journals The Intergenic Small Non-Coding RNA ittA is Required for Optimal Infectivity and Tissue Tropism in Borrelia burgdorferi

2020 ◽  
Author(s):  
Diana N. Medina-Pérez ◽  
Beau Wager ◽  
Erin Troy ◽  
Lihui Gao ◽  
Steven J. Norris ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Borrelia Burgdorferi ◽  
The United States ◽  
Heart Tissue ◽  
Translational Efficiency ◽  
Rna Molecules ◽  
Regulatory Processes ◽  
Non Coding Rna ◽  
Small Regulatory Rnas ◽  
Post Transcriptional Regulation

ABSTRACTPost-transcriptional regulation via small regulatory RNAs (sRNAs) has been implicated in diverse regulatory processes in bacteria, including virulence. One class of sRNAs, termed trans-acting sRNAs, can affect the stability and/or the translational efficiency of regulated transcripts. In this study, we utilized a collaborative approach that employed data from infection with the Borrelia burgdorferi Tn library, coupled with Tn-seq, together with borrelial sRNA and total RNA transcriptomes, to identify an intergenic trans-acting sRNA, which we designate here as ittA for infectivity-associated and tissue-tropic sRNA locus A. The genetic inactivation of ittA resulted in a significant attenuation in infectivity, with decreased spirochetal load in ear, heart, skin and joint tissues. In addition, the ittA mutant did not disseminate to peripheral skin sites or heart tissue, suggesting a role for ittA in regulating a tissue-tropic response. RNA-Seq analysis determined that 19 transcripts were differentially expressed in the ittA mutant relative to its genetic parent, including vraA, bba66, ospD and oms28 (bba74). Subsequent proteomic analyses also showed a significant decrease of OspD and Oms28 (BBA74) proteins. To our knowledge this is the first documented intergenic sRNA that alters the infectivity potential of B. burgdorferi.AUTHOR SUMMARYLyme disease is a tick-borne infection mediated by the spirochetal bacterium, Borrelia burgdorferi, that is responsible for greater than 300,000 infections in the United States per year. As such, additional knowledge regarding how this pathogen modulates its regulatory armamentarium is needed to understand how B. burgdorferi establishes and maintains infection. The identification and characterization of small, non-coding RNA molecules in living systems, designated as sRNAs, has recalibrated how we view post-transcriptional regulation. Recently, over 1,000 sRNAs were identified in B. burgdorferi. Despite the identification of these sRNAs, we do not understand how they affect infectivity or B. burgdorferi pathogenesis related outcomes. Here, we characterize the ittA B. burgdorferi sRNA and show that it is essential for optimal infection using murine experimental infection as our readout. We also track the effect of this sRNA on the transcriptional and proteomic profile as the first step in providing mechanistic insight into how this important sRNA mediates its regulatory effect.

The impact of epitranscriptomic marks on post-transcriptional regulation in plants

2020 ◽  
Author(s):  
Xiang Yu ◽  
Bishwas Sharma ◽  
Brian D Gregory
Keyword(s):  
Transcriptional Regulation ◽  
Plant Development ◽  
Messenger Rna ◽  
Rna Modifications ◽  
Abiotic And Biotic Stresses ◽  
Rna Molecules ◽  
Biotic Stresses ◽  
Specific Mrnas ◽  
Post Transcriptional Regulation ◽  
The Impact

Abstract Ribonucleotides within the various RNA molecules in eukaryotes are marked with more than 160 distinct covalent chemical modifications. These modifications include those that occur internally in messenger RNA (mRNA) molecules such as N6-methyladenosine (m6A) and 5-methylcytosine (m5C), as well as those that occur at the ends of the modified RNAs like the non-canonical 5′ end nicotinamide adenine dinucleotide (NAD+) cap modification of specific mRNAs. Recent findings have revealed that covalent RNA modifications can impact the secondary structure, translatability, functionality, stability and degradation of the RNA molecules in which they are included. Many of these covalent RNA additions have also been found to be dynamically added and removed through writer and eraser complexes, respectively, providing a new layer of epitranscriptome-mediated post-transcriptional regulation that regulates RNA quality and quantity in eukaryotic transcriptomes. Thus, it is not surprising that the regulation of RNA fate mediated by these epitranscriptomic marks has been demonstrated to have widespread effects on plant development and the responses of these organisms to abiotic and biotic stresses. In this review, we highlight recent progress focused on the study of the dynamic nature of these epitranscriptome marks and their roles in post-transcriptional regulation during plant development and response to environmental cues, with an emphasis on the mRNA modifications of non-canonical 5′ end NAD+ capping, m6A and several other internal RNA modifications.

scholarly journals Our First Experience with Magnetic Separation of Platelets for Analyses of Platelet MicroRNA in Patients with Sticky Platelet Syndrome

2019 ◽  
Vol 19 (3) ◽  
pp. 88-94
Author(s):  
L Vadelova ◽  
J Ivankova ◽  
J Sokol ◽  
M Skerenova ◽  
J Zolkova ◽  
...  
Keyword(s):  
Magnetic Separation ◽  
Platelet Reactivity ◽  
Platelet Rich Plasma ◽  
Expression Profiles ◽  
Adenosine Diphosphate ◽  
Blood Samples ◽  
Rna Molecules ◽  
Non Coding Rna ◽  
Low Concentrations ◽  
Post Transcriptional Regulation

Abstract Introduction: Sticky platelet syndrome (SPS) is referred to as a platelet hyperaggregability triggered by low concentrations of platelet agonists adenosine diphosphate (ADP) and/or epinephrine (EPI). Platelet aggregation with other inducers (collagen, arachidonic acid, ristocetin, and thrombin) remains within a normal range. MicroRNAs (miRNAs) are small, non-coding RNA molecules that play an important role in post-transcriptional regulation of protein expression. More recently, several studies show that the platelets are an abundant source of miRNAs and that the miRNA expression profiles within platelets correlate with the platelet reactivity. Aim: The principle objective of this article is to describe the method which we developed for the preparation of the pure platelet samples and report the results of this method. These final pure platelet samples are intended to be the first step for the platelet miRNA testing. Methods: The blood samples from 50 subjects were examined in the study. Then, the platelet rich plasma (PRP) samples obtained by centrifugation of the patient blood samples were used for our experiments. Subsequently, the erythrocytes and leucocytes remaining in PRP sample were magnetically labelled by CD45 Microbeads and CD235a Microbeads. After incubation the PRP sample passed through the magnetic separation system and the magnetically labelled cells (erythrocytes and leucocytes) were retained within the column of separator. The number of cells in the final PRP samples was measured by the blood cell analyser. Results and conclusion: We successfully developed and optimized the effective and reproducible method for magnetic separation of platelets, resulting in the leukocyte-depleted and erythrocyte-depleted platelet samples, which can be used for further genetic analyses.

scholarly journals Poly(A) tail length is a major regulator of maternal gene expression during the mammalian oocyte-to-embryo transition

2021 ◽  
Author(s):  
Yusheng Liu ◽  
Hu Nie ◽  
Chuanxin Zhang ◽  
Zhenzhen Hou ◽  
Jiaqiang Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transcriptional Regulation ◽  
Tail Length ◽  
Human Embryos ◽  
Molecular Evidence ◽  
Translational Efficiency ◽  
Mammalian Oocyte ◽  
Maternal Mrna ◽  
Polyadenylation Sites ◽  
Post Transcriptional Regulation

AbstractTranscription is silent during the mammalian oocyte-to-embryo transition (OET) until zygotic genome activation (ZGA). Therefore, the OET relies on post-transcriptional regulation of maternal mRNA, among which poly(A) tail lengths have been found to regulate translation for a small number of genes1–3. However, transcriptome-wide poly(A) tail length dynamics and their role in gene expression during the mammalian OET remain unknown. Here, we quantified transcriptome-wide mRNA poly(A) tail length dynamics during the mammalian OET using PAIso-seq1 and PAIso-seq24,5, two methods with different underlying principles that preserve the poly(A) tail information. We revealed that poly(A) tail length was highly dynamic during the mouse OET, and Btg4 is responsible for global maternal mRNA deadenylation. We found that the poly(A) tail length positively associated with translational efficiency transcriptome-wide in mouse oocytes. In addition, genes with different alternative polyadenylation isoforms show longer poly(A) tails for isoforms with distal polyadenylation sites compared to those with proximal polyadenylation sites in mouse, rat, pig and human oocytes after meiotic resumption, which is not seen in cultured cell lines. Surprisingly, mammalian embryos, namely mouse, rat, pig, and human embryos, all experience highly conserved global mRNA re-polyadenylation after fertilization, providing molecular evidence that the early embryo development before ZGA is driven by re-polyadenylated maternal mRNAs rather than newly transcribed mRNAs. Together, our study reveals the conserved mRNA poly(A) tail length landscape. This resource can be used for exploring spatiotemporal post-transcriptional regulation throughout the mammalian OET.

scholarly journals The Etiological Agent of Lyme Disease, Borrelia burgdorferi, Appears To Contain Only a Few Small RNA Molecules

2004 ◽  
Vol 186 (24) ◽  
pp. 8472-8477 ◽  
Author(s):  
Yngve Östberg ◽  
Ignas Bunikis ◽  
Sven Bergström ◽  
Jörgen Johansson
Keyword(s):  
Borrelia Burgdorferi ◽  
Small Rna ◽  
Rna Binding ◽  
Rna Binding Protein ◽  
Rnase E ◽  
Regulatory Pathways ◽  
Rna Molecules ◽  
Small Regulatory Rnas ◽  
Regulatory Rnas

ABSTRACT Small regulatory RNAs (sRNAs) have recently been shown to be the main controllers of several regulatory pathways. The function of sRNAs depends in many cases on the RNA-binding protein Hfq, especially for sRNAs with an antisense function. In this study, the genome of Borrelia burgdorferi was subjected to different searches for sRNAs, including direct homology and comparative genomics searches and ortholog- and annotation-based search strategies. Two new sRNAs were found, one of which showed complementarity to the rpoS region, which it possibly controls by an antisense mechanism. The role of the other sRNA is unknown, although observed complementarities against particular mRNA sequences suggest an antisense mechanism. We suggest that the low level of sRNAs observed in B. burgdorferi is at least partly due to the presumed lack of both functional Hfq protein and RNase E activity.

scholarly journals Thioredoxin post-transcriptional regulation by H19 provides a new function to mRNA-like non-coding RNA

Oncogene ◽  
2002 ◽  
Vol 21 (10) ◽  
pp. 1625-1631 ◽  
Author(s):  
Séverine Lottin ◽  
Anne-Sophie Vercoutter-Edouart ◽  
Eric Adriaenssens ◽  
Xavier Czeszak ◽  
Jérôme Lemoine ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Non Coding Rna ◽  
Post Transcriptional Regulation

scholarly journals Severe dysregulation of TNF expression leads to multiple inflammatory diseases and embryonic death

2020 ◽  
Author(s):  
Elise Clayer ◽  
Destiny Dalseno ◽  
Andrew Kueh ◽  
Derek Lacey ◽  
Minhsuang Tsai ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Inflammatory Diseases ◽  
Regulatory Element ◽  
Translational Efficiency ◽  
Rapid Induction ◽  
Regulation Mechanisms ◽  
Post Transcriptional Regulation ◽  
First Time ◽  
Embryonic Death

AbstractPost-transcriptional regulation mechanisms regulate mRNA stability or translational efficiency via ribosomes and recent evidence indicates that it is a major determinant of the accurate levels of cytokine mRNAs. While transcriptional regulation of Tnf has been well studied and found to be important for the rapid induction of Tnf mRNA and regulation of the acute phase of inflammation, study of its post-transcriptional regulation has been largely limited to the role of the AU-rich element (ARE), and to a lesser extent, that of the constitutive decay element (CDE). We have identified a new regulatory element (NRE) in the 3’ untranslated region (3’UTR) of Tnf, and demonstrate that ARE, CDE and NRE cooperate to efficiently down regulate Tnf expression and prevent autoimmune inflammatory diseases. We also show for the first time that excessive TNF may lead to embryonic death.

scholarly journals Spectroscopic observation of RNA chaperone activities of Hfq in post-transcriptional regulation by a small non-coding RNA

2007 ◽  
Vol 35 (3) ◽  
pp. 999-1006 ◽  
Author(s):  
Véronique Arluison ◽  
Sungchul Hohng ◽  
Rahul Roy ◽  
Olivier Pellegrini ◽  
Philippe Régnier ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Spectroscopic Observation ◽  
Rna Chaperone ◽  
Non Coding Rna ◽  
Post Transcriptional Regulation
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