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Molecular and functional characterization of single-box high-mobility group B (HMGB) chromosomal protein from Aedes aegypti
- Source :
- Gene. 671
- Publication Year :
- 2018
-
Abstract
- High-mobility group B (HMGB) proteins have highly conserved, unique DNA-binding domains, HMG boxes, that can bind non-B-type DNA structures, such as bent, kinked and unwound structures, with high affinity. HMGB proteins also promote DNA bending, looping and unwinding. In this study, we determined the role of the Aedes aegypti single HMG-box domain protein AaHMGB; characterized its structure, spatiotemporal expression levels, subcellular localization, and nucleic acid binding activities; and compared these properties with those of its double-HMG-box counterpart protein, AaHMGB1. Via qRT-PCR, we showed that AaHMGB is expressed at much higher levels than AaHMGB1 throughout mosquito development. In situ hybridization results suggested a role for AaHMGB and AaHMGB1 during embryogenesis. Immunolocalization in the midgut revealed that AaHMGB is exclusively nuclear. Circular dichroism and fluorescence spectroscopy analyses showed that AaHMGB exhibits common features of α-helical structures and is more stably folded than AaHMGB1, likely due to the presence of one or two HMG boxes. Using several DNA substrates or single-stranded RNAs as probes, we observed significant differences between AaHMGB and AaHMGB1 in terms of their binding patterns, activity and/or specificity. Importantly, we showed that the phosphorylation of AaHMGB plays a critical role in its DNA-binding activity. Our study provides additional insight into the roles of single- versus double-HMG-box-containing proteins in nucleic acid interactions for better understanding of mosquito development, physiology and homeostasis.
- Subjects :
- 0301 basic medicine
Models, Molecular
Circular dichroism
Protein domain
In situ hybridization
Biology
Protein Structure, Secondary
03 medical and health sciences
chemistry.chemical_compound
Aedes
HMGB Proteins
Genetics
Animals
Tissue Distribution
Phosphorylation
Cell Nucleus
Circular Dichroism
Gene Expression Regulation, Developmental
General Medicine
Subcellular localization
Cell biology
030104 developmental biology
High-mobility group
chemistry
Nucleic acid
Insect Proteins
DNA
Subjects
Details
- ISSN :
- 18790038
- Volume :
- 671
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....1f0df7010afbd5424403826f54eda8e8