In vitro reconstitution of yeast RNA polymerase II transcription initiation with high efficiency

Publication date: Available online 21 March 2019Source: MethodsAuthor(s): Rina Fujiwara, Kenji MurakamiAbstractTranscription initiation can be reconstituted from highly purified general transcription factors (GTFs), RNA polymerase II (pol II), and promoter DNA. However, earlier biochemical reconstitution systems had a serious technical limitation, namely very poor initiation efficiency. Due to the poor efficiency of the reaction and trace amounts of proteins involved in the pre-initiation complex (PIC) assembly, detection of transcription and PIC formation was only possible by the synthesis of a radiolabeled transcript and by immunoblotting for PIC components on templates. Here we describe a transcription system that is capable of initiating transcription with>90% efficiency using homogeneous, active yeast components including TFIIA, TFIIB, TBP, TFIIE, TFIIF, TFIIH, Sub1, and pol II. The abundant specifically assembled PICs on promoter DNA can be separated from free general transcription factors (GTFs) and pol II by density gradient sedimentation, irrespective of the length of promoter DNA. The system is robust, and can be modified to accommodate many other transcription factors, and the resulting complexes can be analyzed by SDS-PAGE followed by Coomassie Blue staining. This technical advance now paves the way to conduct definitive biochemical and structural studies of the complete process of pol II initiation from the PIC, through promoter escape, and finally to producti...
Source: Methods - Category: Molecular Biology Source Type: research

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Publication date: Available online 24 April 2019Source: Journal of Critical CareAuthor(s): Philippe Montravers, Ignacio Martin-Loeches
Source: Journal of Critical Care - Category: Gastroenterology Source Type: research
Publication date: Available online 25 April 2019Source: European Annals of Otorhinolaryngology, Head and Neck DiseasesAuthor(s): G. Dumas, H. Tan, L. Dumas, P. Perrin, A. Lion, S. SchmerberAbstractObjectiveTo establish optimum stimulus frequency and location of bone conducted vibration provoking a skull vibration induced nystagmus (SVIN) in superior semi-circular canal dehiscences.MethodsSVIN 3D components in 40 patients with semi-circular canal dehiscence (27 unilateral and 13 bilateral) were compared with a group of 18 patients with severe unilateral vestibular loss and a control group of 11 volunteers.ResultsIn unilater...
Source: European Annals of Otorhinolaryngology, Head and Neck Diseases - Category: ENT & OMF Source Type: research
Source: Auris, Nasus, Larynx - Category: ENT & OMF Source Type: research
Source: Auris, Nasus, Larynx - Category: ENT & OMF Source Type: research
Conclusion: The high IGF-II/IGFBP-3 molar ratio was not the cause of the sustained high growth velocity, due to the low affinity of IGF-II for IGF type 1 receptor. The key finding was theHESX1 mutation, as similar cases have been described before, suggesting a common mechanism for growth without GH. However, the variable penetrance of this variant in heterozygous relatives suggests that modifier genes or mechanisms involving combinations with mutations of other genes involved in intracellular signaling pathways might be responsible.Horm Res Paediatr
Source: Hormone Research in Paediatrics - Category: Endocrinology Source Type: research
Publication date: Available online 24 April 2019Source: The Lancet Diabetes &EndocrinologyAuthor(s): Stuart W Flint
Source: The Lancet Diabetes and Endocrinology - Category: Endocrinology Source Type: research
Publication date: Available online 24 April 2019Source: The Lancet Diabetes &EndocrinologyAuthor(s): Peter H Bennett
Source: The Lancet Diabetes and Endocrinology - Category: Endocrinology Source Type: research
Publication date: Available online 13 April 2019Source: Molecular and Cellular EndocrinologyAuthor(s): Matti Poutanen, Trevor M. Penning
Source: Molecular and Cellular Endocrinology - Category: Endocrinology Source Type: research
ConclusionSGLT2 inhibitors have relatively safe profiles and can efficiently decrease HbA1c as well as fasting and postprandial glucose levels. Furthermore, SGLT2 inhibitors administrations are not associated with significant hypoglycemic episodes or weight gain. Thus, combination of SGLT2 inhibitors and other less harmful anti-diabetic medicines could be considered if there is no any contraindication.
Source: Diabetes and Metabolic Syndrome: Clinical Research and Reviews - Category: Endocrinology Source Type: research
ConclusionDiabetes was correlated significantly with dyslipidemia and atherogenic risk in parallel with an increase in IFN-γ production and hematological inflammatory biomarkers; leukocytes, neutrophil/lymphocyte and platelet/lymphocyte ratios, MPV and PDW values. The amelioration in inflammatory biomarkers was associated with improvement in glycemic control.
Source: Diabetes and Metabolic Syndrome: Clinical Research and Reviews - Category: Endocrinology Source Type: research
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