The impact of CRISPR repeat sequence on structures of a Cas6 protein-RNA complex
- FSU
The repeat-associated mysterious proteins (RAMPs) comprise the most abundant family of proteins involved in prokaryotic immunity against invading genetic elements conferred by the clustered regularly interspaced short palindromic repeat (CRISPR) system. Cas6 is one of the first characterized RAMP proteins and is a key enzyme required for CRISPR RNA maturation. Despite a strong structural homology with other RAMP proteins that bind hairpin RNA, Cas6 distinctly recognizes single-stranded RNA. Previous structural and biochemical studies show that Cas6 captures the 5' end while cleaving the 3' end of the CRISPR RNA. Here, we describe three structures and complementary biochemical analysis of a noncatalytic Cas6 homolog from Pyrococcus horikoshii bound to CRISPR repeat RNA of different sequences. Our study confirms the specificity of the Cas6 protein for single-stranded RNA and further reveals the importance of the bases at Positions 5-7 in Cas6-RNA interactions. Substitutions of these bases result in structural changes in the protein-RNA complex including its oligomerization state.
- Research Organization:
- Advanced Photon Source (APS), Argonne National Laboratory (ANL), Argonne, IL (US)
- Sponsoring Organization:
- NSFDOE - BASIC ENERGY SCIENCES
- OSTI ID:
- 1034950
- Journal Information:
- Protein Sci., Journal Name: Protein Sci. Journal Issue: (3) ; 03, 2012 Vol. 21
- Country of Publication:
- United States
- Language:
- ENGLISH
Similar Records
Evolution of CRISPR RNA recognition and processing by Cas6 endonucleases
Cas6 is an endoribonuclease that generates guide RNAs for invader defense in prokaryotes
Cas5d Protein Processes Pre-crRNA and Assembles into a Cascade-like Interference Complex in Subtype I-C/Dvulg CRISPR-Cas System
Journal Article
·
Sun Oct 20 20:00:00 EDT 2013
· Nucleic Acids Research
·
OSTI ID:1625531
Cas6 is an endoribonuclease that generates guide RNAs for invader defense in prokaryotes
Journal Article
·
Mon Nov 08 23:00:00 EST 2010
· Gene Dev.
·
OSTI ID:1007160
Cas5d Protein Processes Pre-crRNA and Assembles into a Cascade-like Interference Complex in Subtype I-C/Dvulg CRISPR-Cas System
Journal Article
·
Wed Oct 10 00:00:00 EDT 2012
· Structure
·
OSTI ID:1050084