Crystal structure and initial characterization of a novel archaeal-like Holliday junction-resolving enzyme from Thermus thermophilus phage Tth15-6
(2022) In Acta crystallographica. Section D, Structural biology 78(Pt 2). p.212-227- Abstract
This study describes the production, characterization and structure determination of a novel Holliday junction-resolving enzyme. The enzyme, termed Hjc_15-6, is encoded in the genome of phage Tth15-6, which infects Thermus thermophilus. Hjc_15-6 was heterologously produced in Escherichia coli and high yields of soluble and biologically active recombinant enzyme were obtained in both complex and defined media. Amino-acid sequence and structure comparison suggested that the enzyme belongs to a group of enzymes classified as archaeal Holliday junction-resolving enzymes, which are typically divalent metal ion-binding dimers that are able to cleave X-shaped dsDNA-Holliday junctions (Hjs). The crystal structure of Hjc_15-6 was... (More)
This study describes the production, characterization and structure determination of a novel Holliday junction-resolving enzyme. The enzyme, termed Hjc_15-6, is encoded in the genome of phage Tth15-6, which infects Thermus thermophilus. Hjc_15-6 was heterologously produced in Escherichia coli and high yields of soluble and biologically active recombinant enzyme were obtained in both complex and defined media. Amino-acid sequence and structure comparison suggested that the enzyme belongs to a group of enzymes classified as archaeal Holliday junction-resolving enzymes, which are typically divalent metal ion-binding dimers that are able to cleave X-shaped dsDNA-Holliday junctions (Hjs). The crystal structure of Hjc_15-6 was determined to 2.5 Å resolution using the selenomethionine single-wavelength anomalous dispersion method. To our knowledge, this is the first crystal structure of an Hj-resolving enzyme originating from a bacteriophage that can be classified as an archaeal type of Hj-resolving enzyme. As such, it represents a new fold for Hj-resolving enzymes from phages. Characterization of the structure of Hjc_15-6 suggests that it may form a dimer, or even a homodimer of dimers, and activity studies show endonuclease activity towards Hjs. Furthermore, based on sequence analysis it is proposed that Hjc_15-6 has a three-part catalytic motif corresponding to E-SD-EVK, and this motif may be common among other Hj-resolving enzymes originating from thermophilic bacteriophages.
(Less)
- author
- organization
- publishing date
- 2022-02-01
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Acta crystallographica. Section D, Structural biology
- volume
- 78
- issue
- Pt 2
- pages
- 16 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- scopus:85124061585
- pmid:35102887
- ISSN
- 2059-7983
- DOI
- 10.1107/S2059798321012298
- language
- English
- LU publication?
- yes
- additional info
- open access.
- id
- bbaebebf-e6ad-4d05-82a0-6dd075fe5fda
- date added to LUP
- 2022-02-10 10:06:49
- date last changed
- 2024-04-23 09:34:21
@article{bbaebebf-e6ad-4d05-82a0-6dd075fe5fda, abstract = {{<p>This study describes the production, characterization and structure determination of a novel Holliday junction-resolving enzyme. The enzyme, termed Hjc_15-6, is encoded in the genome of phage Tth15-6, which infects <i>Thermus thermophilus</i>. Hjc_15-6 was heterologously produced in <i>Escherichia coli</i> and high yields of soluble and biologically active recombinant enzyme were obtained in both complex and defined media. Amino-acid sequence and structure comparison suggested that the enzyme belongs to a group of enzymes classified as archaeal Holliday junction-resolving enzymes, which are typically divalent metal ion-binding dimers that are able to cleave X-shaped dsDNA-Holliday junctions (Hjs). The crystal structure of Hjc_15-6 was determined to 2.5 Å resolution using the selenomethionine single-wavelength anomalous dispersion method. To our knowledge, this is the first crystal structure of an Hj-resolving enzyme originating from a bacteriophage that can be classified as an archaeal type of Hj-resolving enzyme. As such, it represents a new fold for Hj-resolving enzymes from phages. Characterization of the structure of Hjc_15-6 suggests that it may form a dimer, or even a homodimer of dimers, and activity studies show endonuclease activity towards Hjs. Furthermore, based on sequence analysis it is proposed that Hjc_15-6 has a three-part catalytic motif corresponding to E-SD-EVK, and this motif may be common among other Hj-resolving enzymes originating from thermophilic bacteriophages.</p>}}, author = {{Ahlqvist, Josefin and Linares-Pastén, Javier A and Håkansson, Maria and Jasilionis, Andrius and Kwiatkowska-Semrau, Karolina and Friðjónsson, Ólafur H. and Kaczorowska, Anna-Karina and Dabrowski, Slawomir and Ævarsson, Arnþór and Hreggviðsson, Guðmundur Ó. and Al-Karadaghi, Salam and Kaczorowski, Tadeusz and Nordberg Karlsson, Eva}}, issn = {{2059-7983}}, language = {{eng}}, month = {{02}}, number = {{Pt 2}}, pages = {{212--227}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Acta crystallographica. Section D, Structural biology}}, title = {{Crystal structure and initial characterization of a novel archaeal-like Holliday junction-resolving enzyme from <i>Thermus thermophilus</i> phage Tth15-6}}, url = {{http://dx.doi.org/10.1107/S2059798321012298}}, doi = {{10.1107/S2059798321012298}}, volume = {{78}}, year = {{2022}}, }