Next Article in Journal
Mitochondrial Pyruvate Carrier Function in Health and Disease across the Lifespan
Next Article in Special Issue
G-Quadruplexes in the Archaea Domain
Previous Article in Journal
Single and Combined Methods to Specifically or Bulk-Purify RNA–Protein Complexes
Previous Article in Special Issue
Regulated Iron Siderophore Production of the Halophilic Archaeon Haloferax volcanii
Open AccessFeature PaperArticle

The CARF Protein MM_0565 Affects Transcription of the Casposon-Encoded cas1-solo Gene in Methanosarcina mazei Gö1

1
Institute of General Microbiology, Christian-Albrechts-University of Kiel, 24118 Kiel, Germany
2
Core Unit Systems Medicine, Institute of Molecular Infection Biology, University of Würzburg, 97070 Würzburg, Germany
3
ZB MED–Information Centre for Life Sciences, 50931 Cologne, Germany
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Biomolecules 2020, 10(8), 1161; https://doi.org/10.3390/biom10081161
Received: 29 May 2020 / Revised: 4 August 2020 / Accepted: 4 August 2020 / Published: 7 August 2020
(This article belongs to the Special Issue Archaea: Diversity, Metabolism and Molecular Biology)
Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) loci are found in bacterial and archaeal genomes where they provide the molecular machinery for acquisition of immunity against foreign DNA. In addition to the cas genes fundamentally required for CRISPR activity, a second class of genes is associated with the CRISPR loci, of which many have no reported function in CRISPR-mediated immunity. Here, we characterize MM_0565 associated to the type I-B CRISPR-locus of Methanosarcina mazei Gö1. We show that purified MM_0565 composed of a CRISPR-Cas Associated Rossmann Fold (CARF) and a winged helix-turn-helix domain forms a dimer in solution; in vivo, the dimeric MM_0565 is strongly stabilized under high salt stress. While direct effects on CRISPR-Cas transcription were not detected by genetic approaches, specific binding of MM_0565 to the leader region of both CRISPR-Cas systems was observed by microscale thermophoresis and electromobility shift assays. Moreover, overexpression of MM_0565 strongly induced transcription of the cas1-solo gene located in the recently reported casposon, the gene product of which shows high similarity to classical Cas1 proteins. Based on our findings, and taking the absence of the expressed CRISPR locus-encoded Cas1 protein into account, we hypothesize that MM_0565 might modulate the activity of the CRISPR systems on different levels. View Full-Text
Keywords: methanoarchaea; CRISPR-Cas system; transcriptional regulation; adaptation phase; casposon; Methanosarcina mazei methanoarchaea; CRISPR-Cas system; transcriptional regulation; adaptation phase; casposon; Methanosarcina mazei
Show Figures

Figure 1

MDPI and ACS Style

Ulbricht, A.; Nickel, L.; Weidenbach, K.; Vargas Gebauer, H.; Kießling, C.; Förstner, K.U.; Schmitz, R.A. The CARF Protein MM_0565 Affects Transcription of the Casposon-Encoded cas1-solo Gene in Methanosarcina mazei Gö1. Biomolecules 2020, 10, 1161.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop