DNA looping by two 5-methylcytosine-binding proteins quantified using nanofluidic devices.
Epigenetics Chromatin
; 13(1): 18, 2020 03 16.
Article
in En
| MEDLINE
| ID: mdl-32178718
BACKGROUND: MeCP2 and MBD2 are members of a family of proteins that possess a domain that selectively binds 5-methylcytosine in a CpG context. Members of the family interact with other proteins to modulate DNA packing. Stretching of DNA-protein complexes in nanofluidic channels with a cross-section of a few persistence lengths allows us to probe the degree of compaction by proteins. RESULTS: We demonstrate DNA compaction by MeCP2 while MBD2 does not affect DNA configuration. By using atomic force microscopy (AFM), we determined that the mechanism for compaction by MeCP2 is the formation of bridges between distant DNA stretches and the formation of loops. CONCLUSIONS: Despite sharing a similar specific DNA-binding domain, the impact of full-length 5-methylcytosine-binding proteins can vary drastically between strong compaction of DNA and no discernable large-scale impact of protein binding. We demonstrate that ATTO 565-labeled MBD2 is a good candidate as a staining agent for epigenetic mapping.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA
/
DNA Methylation
/
5-Methylcytosine
/
Microfluidics
/
DNA-Binding Proteins
/
Methyl-CpG-Binding Protein 2
Limits:
Humans
Language:
En
Journal:
Epigenetics Chromatin
Year:
2020
Document type:
Article
Affiliation country:
United States
Country of publication:
United kingdom