Defining the crucial domain and amino acid residues in bacterial lon protease for DNA binding and processing of DNA-interacting substrates

Anna Karłowicz , Katarzyna Węgrzyn , Marta Gross , Dagmara Kaczyńska , Małgorzata Ropelewska , Małgorzata Siemiątkowska , Janusz M. Bujnicki , Igor Konieczny

Abstract

Lon protease has previously been shown to interact with DNA but the role of this interaction for Lon proteolytic activity has not been characterized. In this study, we used truncated Escherichia Lon constructs, bioinformatics analysis and site-directed mutagenesis to identify Lon domains and residues crucial for Lon binding with DNA and effects on Lon proteolytic activity. We found that deletion of Lon’s ATPase domain abrogated interactions with DNA. Substitution of positively charged amino acids in this domain in full-length Lon with residues conferring a net negative charge disrupted binding of Lon to DNA. These changes also affected the degradation of nucleic acid–binding protein substrates of Lon, intracellular localization of Lon, and cell morphology. In vivo tests revealed that Lon-DNA interactions are essential for Lon activity in cell division control. In summary, we demonstrate that the ability of Lon to bind DNA is determined by its ATPase domain, that this binding is required for processing protein substrates in nucleoprotein complexes, and that Lon may help regulate DNA replication in response to growth conditions.
Author Anna Karłowicz KBMiK
Anna Karłowicz,,
- Department of Molecular and Cellular Biology
, Katarzyna Węgrzyn KBMiK
Katarzyna Węgrzyn,,
- Department of Molecular and Cellular Biology
, Marta Gross MWB UG i GUMed
Marta Gross,,
- Intercollegiate Faculty of Biotechnology UG
, Dagmara Kaczyńska
Dagmara Kaczyńska,,
-
, Małgorzata Ropelewska MWB UG i GUMed
Małgorzata Ropelewska,,
- Intercollegiate Faculty of Biotechnology UG
, Małgorzata Siemiątkowska
Małgorzata Siemiątkowska,,
-
, Janusz M. Bujnicki
Janusz M. Bujnicki,,
-
, Igor Konieczny KBMiK
Igor Konieczny,,
- Department of Molecular and Cellular Biology
Journal seriesJournal of Biological Chemistry, ISSN 0021-9258
Issue year2017
Vol292
No18
Pages7507-7518
Publication size in sheets0.55
Keywords in Englishproteolysis, protein-DNA interaction, DNA binding protein, ATP-dependent protease, Escherichia coli
DOIDOI:10.1074/jbc.M116.766709
URL http://www.jbc.org/content/early/2017/03/14/jbc.M116.766709.full.pdf
Languageen angielski
Score (nominal)35
ScoreMinisterial score = 35.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 35.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 4.125 (2) - 2016=4.323 (5)
Citation count*0
Cite
Share Share



* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
Back