title: Deciphering the role of the AT-rich interaction domain and the HMG-box domain of ARID-HMG proteins of Arabidopsis thaliana.
abstract: ARID-HMG DNA-binding proteins represent a novel group of HMG-box containing protein family where the AT-rich interaction domain (ARID) is fused with the HMG-box domain in a single polypeptide chain. ARID-HMG proteins are highly plant specific with homologs found both in flowering plants as well as in moss such as Physcomitrella. The expression of these proteins is ubiquitous in plant tissues and primarily localises in the cell nucleus. HMGB proteins are involved in several nuclear processes, but the role of ARID-HMG proteins in plants remains poorly explored. Here, we performed DNA-protein interaction studies with Arabidopsis ARID-HMG protein HMGB11 (At1g55650) to understand the functionality of this protein and its individual domains. DNA binding assays revealed that AtHMGB11 can bind double-stranded DNA with a weaker affinity (Kd = 475 ± 17.9 nM) compared to Arabidopsis HMGB1 protein (Kd = 39.8 ± 2.68 nM). AtHMGB11 also prefers AT-rich DNA as a substrate and shows structural bias for supercoiled DNA. Molecular docking of the DNA-AtHMGB11 complex indicated that the protein interacts with the DNA major groove, mainly through its ARID domain and the junction region connecting the ARID and the HMG-box domain. Also, predicted by the docking model, mutation of Lys(85) from the ARID domain and Arg(199) & Lys(202) from the junction region affects the DNA binding affinity of AtHMGB11. In addition, AtHMGB11 and its truncated form containing the HMG-box domain can not only promote DNA mini-circle formation but are also capable of inducing negative supercoils into relaxed plasmid DNA suggesting the involvement of this protein in several nuclear events. Overall, the study signifies that both the ARID and the HMG-box domain contribute to the optimal functioning of ARID-HMG protein in vivo.
authors: Division of Plant Biology, Bose Institute, Kolkata, 700054, India.
authors: Adrita
authors: A
authors: Roy
authors: Department of Chemistry, Bose Institute, Kolkata, 700054, India.
authors: Arkajyoti
authors: A
authors: Dutta
authors: Division of Plant Biology, Bose Institute, Kolkata, 700054, India.
authors: Dipan
authors: D
authors: Roy
authors: Division of Plant Biology, Bose Institute, Kolkata, 700054, India.
authors: Payel
authors: P
authors: Ganguly
authors: School of Biotechnology, Yeungnam University, Gyeongsan, 712-749, South Korea.
authors: Ritesh
authors: R
authors: Ghosh
authors: Department of Biophysics, Bose Institute, Kolkata, 700054, India.
authors: Rajiv K
authors: RK
authors: Kar
authors: Department of Biophysics, Bose Institute, Kolkata, 700054, India.
authors: Anirban
authors: A
authors: Bhunia
authors: Department of Chemistry, Bose Institute, Kolkata, 700054, India.
authors: Jayanta
authors: J
authors: Mukhopadhyay
authors: Division of Plant Biology, Bose Institute, Kolkata, 700054, India. shubho@jcbose.ac.in.
authors: Shubho
authors: S
authors: Chaudhuri
keywords: ARID-HMG; Arabidopsis thaliana; DNA bending; DNA binding; DNA supercoiling; HMG-box protein