BEGIN:VCALENDAR VERSION:2.0 PRODID:-//wp-events-plugin.com//7.2.2.1//EN TZID:Asia/Jerusalem X-WR-TIMEZONE:Asia/Jerusalem BEGIN:VEVENT UID:1159@biology.technion.ac.il DTSTART;TZID=Asia/Jerusalem:20230215T130000 DTEND;TZID=Asia/Jerusalem:20230215T133000 DTSTAMP:20230124T124120Z URL:https://biology.technion.ac.il/en/seminars/msc-graduate-seminar-muhamm ad-makhzumy/ SUMMARY:MSc Graduate Seminar- Muhammad Makhzumy [No Categories] DESCRIPTION:Location: hybrid- in the Faculty Auditorium/ZOOM: https://techn ion.zoom.us/https://technion.zoom.us/j/6141638094 Muhammad Makhzumy\n Af filiation: \n Host:Dr. Shiber Ayala\n Developing a single molecule approac h to selective ribosome profiling\n\n \;\n\nmRNA-protein interactions are at the center of the translation process. Ribosome-associated factors\ , from modifying enzymes to molecular chaperones\, play a pivotal role in facilitating the folding of the emerging nascent chain\, protecting it fro m aberrant interactions in the crowded cytoplasm. However\, the interplay between the various ribosome-associated factors remains largely obscure. W hat is the mode of interplay? Do they compete for binding? Is there a coor dinated handover? By only observing population averages we can miss crucia l mechanistic features. To address this\, we are developing a single molec ule approach\, targeting two canonical ribosome-associated chaperones:  R AC-SSB1  and NAC-alpha (Stress-Seventy subfamily B subunit of the ribosom e-associated complex and Alpha subunit of the Nascent Polypeptide-Associat ed Complex\, respectively). We have generated two chimeric constructs unde r an inducible promoter\; Fusing the co-translationally acting factor RAC- SSB1 with N6-adenosine methyltransferase IME4\, and the co-translationally acting factor NAC-alpha with RNA-specific adenosine deaminase ADAR2 catal ytic domain. Recent development in Oxford nanopore sequencing platform sen sitivity has allowed for the identification of enzymatically modified nucl eotides. We are utilizing this advance to detect co-translational interact ion in vivo\, by labeling the mRNAs nucleotides in close proximity to thos e interactions. Using immunoprecipitation\, we demonstrated that our chime ric constructs show a strong ribosomal association.  Polysome profiling d emonstrated that the attached RNA-modifying enzymes are in proximity to tr anslated mRNA. RNA extraction following induction and sequencing of the en tire orfs using NanoPore revealed significant changes in the electrical ou tput of samples\, strongly suggesting the presence of modified bases in th e test strain. Further analysis will reveal the interplay between SSB1 and NACα and establish a platform for analysis of various co-translational f actors interaction in single-molecule resolution.\n\nThe lecture will be g iven in English LOCATION:hybrid- in the Faculty Auditorium/ZOOM: https://technion.zoom.us/h ttps://technion.zoom.us/j/6141638094 END:VEVENT BEGIN:VTIMEZONE TZID:Asia/Jerusalem X-LIC-LOCATION:Asia/Jerusalem BEGIN:STANDARD DTSTART:20221030T010000 TZOFFSETFROM:+0300 TZOFFSETTO:+0200 TZNAME:IST END:STANDARD END:VTIMEZONE END:VCALENDAR