Variations in the enzymatic activity of S1-type nucleases results from differences in their active site structures

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60077344%3A_____%2F23%3A00584258" target="_blank" >RIV/60077344:_____/23:00584258 - isvavai.cz</a>

Result on the web

<a href="https://www.sciencedirect.com/science/article/pii/S0304416523001228?via%3Dihub" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0304416523001228?via%3Dihub</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.bbagen.2023.130424" target="_blank" >10.1016/j.bbagen.2023.130424</a>

Result language

angličtina

Original language name

Variations in the enzymatic activity of S1-type nucleases results from differences in their active site structures

Original language description

Background: S1-like nucleases are widespread enzymes commonly used in biotechnology and molecular biology. Although it is commonly believed that they are mainly Zn2+ dependent acidic enzymes, we have found that numerous members of this family deviate from this rule. Therefore, in this work, we decided to check how broad is the range of non-zinc-dependent S1-like nucleases and what is the molecular basis of their activities. Methods: S1-like nucleases chosen for analysis were achieved through heterologous expression in appropriate eukaryotic hosts. To characterize nucleases active-site properties, point mutations were introduced in selected positions. The enzymatic activities of wild-type and mutant nucleases were tested by in-gel nuclease activity assay. Results: We discovered that S1-like nucleases encoded by non-vascular plants and single-celled protozoa, like their higher plant homologues, exhibit a large variety of catalytic properties. We have shown that these individual properties are determined by specific non-conserved active site residues. Conclusions: Our findings demonstrate that mutations that occur during evolution can significantly alter the catalytic properties of S1-like nucleases. As a result, different ions can compete for particular S1-type nucleases active sites. This phenomenon undermines the existing classification of S1-like nucleases. General significance: Our findings have numerous implications for applications and understanding the S1-like nucleases biological functions. For example, new biotechnological applications should take into account their unexpected catalytic properties. Moreover, these results demonstrate that the trinuclear zinc-based model commonly used to characterize the catalytic activities of S1-like nucleases is insufficient to explain the actions of non-zinc-dependent members of this family.

Czech name

—

Czech description

—

Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10608 - Biochemistry and molecular biology

Project

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2023

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Biochimica et Biophysica Acta-General Subjects

ISSN

0304-4165

e-ISSN

1872-8006

Volume of the periodical

1867

Issue of the periodical within the volume

10

Country of publishing house

NL - THE KINGDOM OF THE NETHERLANDS

Number of pages

9

Pages from-to

130424

UT code for WoS article

001058918800001

EID of the result in the Scopus database

2-s2.0-85165714520