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Characterization of the zinc finger μ-protein HVO_0758 from Haloferax volcanii : biological roles, zinc binding, and NMR solution structure.
- Source :
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Frontiers in microbiology [Front Microbiol] 2023 Nov 29; Vol. 14, pp. 1280972. Date of Electronic Publication: 2023 Nov 29 (Print Publication: 2023). - Publication Year :
- 2023
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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 Haloferax volcanii 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&#95;2753, was thoroughly characterized in a previous study. In the present study an in-depth analysis of a second protein, HVO&#95;0758, was performed. To achieve this goal, the HVO&#95;0758 protein was produced heterologously in Escherichia coli and homologously in H. volcanii . The purified protein was characterized using various biochemical approaches and NMR spectroscopy. The findings demonstrated that HVO&#95;0758 is indeed a bona fide zinc finger protein, and that all four cysteine residues are essential for folding. The NMR solution structure was solved, revealing that HVO&#95;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 HVO&#95;0758 deletion mutant and, for comparison, the HVO&#95;2753 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 H. volcanii zinc-finger μ-proteins HVO&#95;0758 and HVO&#95;2753 showed many differences. Taken together, two zinc finger μ-proteins of H. volcanii have been characterized intensively, which emerged as pivotal contributors to swarming behavior and biofilm formation.<br />Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.<br /> (Copyright © 2023 Üresin, Pyper, Borst, Hadjeras, Gelhausen, Backofen, Sharma, Schwalbe and Soppa.)
Details
- Language :
- English
- ISSN :
- 1664-302X
- Volume :
- 14
- Database :
- MEDLINE
- Journal :
- Frontiers in microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 38094630
- Full Text :
- https://doi.org/10.3389/fmicb.2023.1280972