8Q5B

Characterization of the zinc finger u-protein HVO_0758 from Haloferax volcanii: biological roles, zinc binding, and NMR solution structure

Summary for 8Q5B

• Entry DOI: 10.2210/pdb8q5b/pdb
• NMR Information: BMRB: 34844
• Descriptor: Small CPxCG-related zinc finger protein (1 entity in total)
• Functional Keywords: zinc-finger, metal binding protein
• Biological source: Haloferax volcanii DS2
• Total number of polymer chains: 1
• Total formula weight: 6495.41

Authors

Pyper, D.J.,Schwalbe, H. (deposition date: 2023-08-08, release date: 2023-12-27)

Primary citation

Uresin, D.,Pyper, D.J.,Borst, A.,Hadjeras, L.,Gelhausen, R.,Backofen, R.,Sharma, C.,Schwalbe, H.,Soppa, J.
Characterization of the zinc finger mu-protein HVO_0758 from Haloferax volcanii : biological roles, zinc binding, and NMR solution structure.
Front Microbiol, 14:1280972-1280972, 2023

PubMed Abstract: It is increasingly recognized that very small proteins (μ-proteins) are ubiquitously found in all species of the three domains of life, and that they fulfill important functions. The halophilic archaeon contains 282 μ-proteins of less than 70 amino acids. Notably, 43 of these contain two C(P)XCG motifs, suggesting their potential to complex a zinc ion. To explore the significance of these proteins, 16 genes encoding C(P)XCG proteins had been deleted, and the majority of mutants exhibited phenotypic differences to the wild-type. One such protein, HVO_2753, was thoroughly characterized in a previous study. In the present study an in-depth analysis of a second protein, HVO_0758, was performed. To achieve this goal, the HVO_0758 protein was produced heterologously in and homologously in . The purified protein was characterized using various biochemical approaches and NMR spectroscopy. The findings demonstrated that HVO_0758 is indeed a zinc finger protein, and that all four cysteine residues are essential for folding. The NMR solution structure was solved, revealing that HVO_0758 is comprised of an N-terminal alpha helix containing several positively charged residues and a globular core with the zinc finger domain. The transcriptomes of the deletion mutant and, for comparison, the deletion mutant were analyzed with RNA-Seq and compared against that of the wild-type. In both mutants many motility and chemotaxis genes were down-regulated, in agreement to the phenotype of the deletion mutants, which had a swarming deficit. The two zinc-finger μ-proteins HVO_0758 and HVO_2753 showed many differences. Taken together, two zinc finger μ-proteins of have been characterized intensively, which emerged as pivotal contributors to swarming behavior and biofilm formation.

PubMed: 38094630
DOI: 10.3389/fmicb.2023.1280972

Experimental method

SOLUTION NMR