GTP interacts with the γ-subunit of eukaryotic initiation factor eIF-2

AbstractEukaryotic initiation factor eIF-2 is an oligomeric protein consisting of three different subunits. During initiation of protein synthesis eIF-2 interacts with GTP, Met-tRNAf and 40 S ribosomal subunit. By affinity labeling with a photo-reactive GTP analogue it was shown that in the binary complex [eIF-2 · GTP] GTP is in contact with the γ-subunit of eIF-2

Affinity labelling of Escherichia coli ribosomes with a benzylidene derivative of AUGU6 within initiation and pretranslocational complexes

AbstractAffinity labelling of E. coli ribosomes with the 2',3'-O-[4-(N-2-chloroethyl)-N-methylamino]benzylidene derivative of AUGU6 was studied within the initiation complex (complex I) obtained by using fMet-tRNAfMet and initiation factors and within the pretranslocational complex (complex II) obtained by treatment of complex I with the ternary complex Phe-tRNAPhe-GTP-EF-Tu. Both proteins and rRNA of 30 S as well as 50 S subunits were found to be labelled. Sets of proteins labelled within complexes I and II differ considerably. Within complex II, proteins S13 and L10 were labelled preferentially. On the other hand, within complex I, multiple modification is observed (proteins S4, S12, S13, S14, S15, S18, S19, S20/L26 were found to be alkylated) despite the single fixation of a template in the ribosome by interaction of the AUG codon with fMet-tRNAfMet

WIF-1 gene methylation in cervical squamous intraepithelial lesions

Objective. To evaluate the diagnostic value of WIF-1 gene promoter region methylation in the development of cervical intraepithelial neoplasia. Subjects and methods. The investigation included 62 patients aged 18 to 55 years who had come to the Research Center of Obstetrics, Gynecology, and Perinatology, Ministry of Health of the Russian Federation, in the period February to August 2016 for examination and undergone liquid-based cytology, quantitative and qualitative tests for human papillomavirus (HPV), histological examination of cervical biopsy specimens, and extended colposcopy. Bisulfite sequencing was used to study the level of WIF-1 promoter methylation in 62 samples of cells taken from the cervical canal with a cervix brush. Results. The normal mean level of WIF-1 promoter region methylation was 2.3±5.4%. The women diagnosed with high-or low-grade squamous intraepithelial lesions were observed to have a more statistically significant than normal value (p < 0.0001), abnormal WIF-1 gene promoter region hypermethylation at mean frequencies of 29.2±17.2 and 54.8±18.7%, respectively. Conclusion. The findings suggest that the level of WIF-1 gene promoter region methylation significantly correlates with the stage of HPV-associated cervical disease. Thus, evaluation of the WIF-1 gene methylation status may be regarded as a potential diagnostic marker for cervical carcinogenesis and a predictive clinical marker during combination treatment for cervical cancer. © 2017, Bionika Media Ltd. All rights reserved

WIF-1 gene methylation in cervical squamous intraepithelial lesions

Objective. To evaluate the diagnostic value of WIF-1 gene promoter region methylation in the development of cervical intraepithelial neoplasia. Subjects and methods. The investigation included 62 patients aged 18 to 55 years who had come to the Research Center of Obstetrics, Gynecology, and Perinatology, Ministry of Health of the Russian Federation, in the period February to August 2016 for examination and undergone liquid-based cytology, quantitative and qualitative tests for human papillomavirus (HPV), histological examination of cervical biopsy specimens, and extended colposcopy. Bisulfite sequencing was used to study the level of WIF-1 promoter methylation in 62 samples of cells taken from the cervical canal with a cervix brush. Results. The normal mean level of WIF-1 promoter region methylation was 2.3±5.4%. The women diagnosed with high-or low-grade squamous intraepithelial lesions were observed to have a more statistically significant than normal value (p < 0.0001), abnormal WIF-1 gene promoter region hypermethylation at mean frequencies of 29.2±17.2 and 54.8±18.7%, respectively. Conclusion. The findings suggest that the level of WIF-1 gene promoter region methylation significantly correlates with the stage of HPV-associated cervical disease. Thus, evaluation of the WIF-1 gene methylation status may be regarded as a potential diagnostic marker for cervical carcinogenesis and a predictive clinical marker during combination treatment for cervical cancer. © 2017, Bionika Media Ltd. All rights reserved