A Single-Molecule View of Archaeal Transcription. Issue 20 (20th September 2019)

• Record Type: Journal Article
• Authors: Kramm, Kevin; Endesfelder, Ulrike; Grohmann, Dina
• Is Part Of: Journal of molecular biology. Volume 431:Issue 20(2019)
• Page Start: 4116
• Page End: 4131
• Publication Date: 2019-09-20
• Subjects: Archaea -- single-molecule FRET -- nano-positioning system -- single-molecule localization microscopy -- RNA polymerase; RNAP RNA polymerase -- SMLM single-molecule localization microscopy -- SMT single-molecule tracking -- BRE B recognition element -- TBP TATA-binding protein -- TFB transcription factor B -- IC initiation complex -- EC elongation complex -- NTS non-template strand -- NPS nano-positioning system -- PIC pre-initiation complex; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.06.009
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: The discovery of the archaeal domain of life is tightly connected to an in-depth analysis of the prokaryotic RNA world. In addition to Carl Woese's approach to use the sequence of the 16S rRNA gene as phylogenetic marker, the finding of Karl Stetter and Wolfram Zillig that archaeal RNA polymerases (RNAPs) were nothing like the bacterial RNAP but are more complex enzymes that resemble the eukaryotic RNAPII was one of the key findings supporting the idea that archaea constitute the third major branch on the tree of life. This breakthrough in transcriptional research 40 years ago paved the way for in-depth studies of the transcription machinery in archaea. However, although the archaeal RNAP and the basal transcription factors that fine-tune the activity of the RNAP during the transcription cycle are long known, we still lack information concerning the architecture and dynamics of archaeal transcription complexes. In this context, single-molecule measurements were instrumental as they provided crucial insights into the process of transcription initiation, the architecture of the initiation complex and the dynamics of mobile elements of the RNAP. In this review, we discuss single-molecule approaches suitable to examine molecular mechanisms of transcription and highlight findings that shaped our understanding of the archaeal transcription apparatus. We furthermore explore the possibilities and challenges of next-generation single-molecule techniques, for example, super-resolution microscopy and single-molecule tracking, and ask whether these approaches will ultimately allow us to investigate archaeal transcription in vivo . Graphical Abstract: Unlabelled Image Highlights: Single-molecule FRET reveals the dynamics of archaeal transcription initiation. The clamp domain of the archaeal RNAP adopts different conformational states. The architecture of the archaeal and eukaryotic initiation complex differs. Super-resolution microscopy remains challenging in archaea.