Journal
BIOCHEMISTRY
Volume 53, Issue 37, Pages 5848-5863Publisher
AMER CHEMICAL SOC
DOI: 10.1021/bi500470b
Keywords
-
Categories
Funding
- National Science Council [100-2113-M-001-031-MY2, 101-2627-M-001-004]
- Academia Sinica, Taiwan
- Association for International Cancer Research (AICR Project) [10-0570]
- Cancer Research UK
- Wellcome Trust
- International Human Frontier Science Program [CDA-00025/2010-C]
- Biotechnology and Biological Sciences Research Council (BBSRC)
- Medical Research Council (MRC)
- Biotechnology and Biological Sciences Research Council [977500] Funding Source: researchfish
- Worldwide Cancer Research [10-0570] Funding Source: researchfish
Ask authors/readers for more resources
Noncoding Y RNAs are small stem-loop RNAs that are involved in different cellular processes, including the regulation of DNA replication. An evolutionarily conserved small domain in the upper stem of vertebrate Y RNAs has an essential function for the initiation of chromosomal DNA replication. Here we provide a structure-function analysis of this essential RNA domain under physiological conditions. Solution state nuclear magnetic resonance and far-ultraviolet circular dichroism spectroscopy show that the upper stem domain of human Y1 RNA adopts a locally destabilized A-form helical structure involving eight Watson-Crick base pairs. Within this helix, two G:C base pairs are highly stable even at elevated temperatures and therefore may serve as clamps to maintain the local structure of the helix. These two stable G:C base pairs frame three unstable base pairs, which are located centrally between them. Systematic substitution mutagenesis results in a disruption of the ordered A-form helical structure and in the loss of DNA replication initiation activity, establishing a positive correlation between folding stability and function. Our data thus provide a structural basis for the evolutionary conservation of key nucleotides in this RNA domain that are essential for the functionality of noncoding Y RNAs during the initiation of DNA replication.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available