Characterization of the Chimeric PriB-SSBc Protein

doi:10.3390/ijms221910854

中山醫學大學機構典藏 CSMUIR > 中山醫學大學研究成果 > 期刊論文 > Item 310902500/23856

Please use this identifier to cite or link to this item: https://ir.csmu.edu.tw:8080/ir/handle/310902500/23856

Title:	Characterization of the Chimeric PriB-SSBc Protein
Authors:	Lin, ES;Huang, YH;Huang, CY
Keywords:	PriB;SSB;PriA;replication fork;PriB-SSBc;DnaT;primosome;OB fold;DNA mimic;GGRQ motif
Date:	2021
Issue Date:	2022-08-05T09:43:59Z (UTC)
Publisher:	MDPI
Abstract:	PriB is a primosomal protein required for the replication fork restart in bacteria. Although PriB shares structural similarity with SSB, they bind ssDNA differently. SSB consists of an N-terminal ssDNA-binding/oligomerization domain (SSBn) and a flexible C-terminal protein-protein interaction domain (SSBc). Apparently, the largest difference in structure between PriB and SSB is the lack of SSBc in PriB. In this study, we produced the chimeric PriB-SSBc protein in which Klebsiella pneumoniae PriB (KpPriB) was fused with SSBc of K. pneumoniae SSB (KpSSB) to characterize the possible SSBc effects on PriB function. The crystal structure of KpSSB was solved at a resolution of 2.3 angstrom (PDB entry 7F2N) and revealed a novel 114-GGRQ-117 motif in SSBc that pre-occupies and interacts with the ssDNA-binding sites (Asn14, Lys74, and Gln77) in SSBn. As compared with the ssDNA-binding properties of KpPriB, KpSSB, and PriB-SSBc, we observed that SSBc could significantly enhance the ssDNA-binding affinity of PriB, change the binding behavior, and further stimulate the PriA activity (an initiator protein in the pre-primosomal step of DNA replication), but not the oligomerization state, of PriB. Based on these experimental results, we discuss reasons why the properties of PriB can be retrofitted when fusing with SSBc.
URI:	http://dx.doi.org/10.3390/ijms221910854 https://www.webofscience.com/wos/woscc/full-record/WOS:000708039100001 https://ir.csmu.edu.tw:8080/handle/310902500/23856
Relation:	INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES ,2021,v22,issue 19
Appears in Collections:	[中山醫學大學研究成果] 期刊論文

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