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Citation for Study 24443

Citation title: "Arabidopsis thaliana organelles mimic the T7 phage DNA replisome with specific interactions between Twinkle protein and DNA polymerases Pol1A and Pol1B".

Study name: "Arabidopsis thaliana organelles mimic the T7 phage DNA replisome with specific interactions between Twinkle protein and DNA polymerases Pol1A and Pol1B".

This study is part of submission 24443 (Status: Published).

Citation

Morley S., Peralta-castro A., Brieba L.G., Miller J., Ong K., Ridge P., Oliphant A., Aldous S., & Nielsen B. 2019. Arabidopsis thaliana organelles mimic the T7 phage DNA replisome with specific interactions between Twinkle protein and DNA polymerases Pol1A and Pol1B. BMC Plant Biology, .

Authors

• Morley S. (submitter)
• Peralta-castro A.
• Brieba L.G.
• Miller J.
• Ong K.
• Ridge P.
• Oliphant A.
• Aldous S.
• Nielsen B. 8014221102

Abstract

Background Plant chloroplasts and mitochondria utilize nuclear encoded proteins to replicate their DNA. These proteins are purposely built for replication in the organelle environment and are distinct from those involved in replication of the nuclear genome. These organelle-localized proteins also have ancestral roots in bacterial and bacteriophage genes, supporting the endosymbiotic theory of their origin. We examined the interactions between three of these proteins from Arabidopsis thaliana: a DNA helicase-primase similar to bacteriophage T7 gp4 protein and animal mitochondrial Twinkle, and two DNA polymerases, Pol1A and Pol1B. We used a three- pronged approach to analyze the interactions, including Yeast-two-hybrid analysis, Direct Coupling Analysis (DCA), and thermophoresis. Results Yeast-two-Hybrid analysis reveals residues 120-295 of Twinkle as the minimal region that can still interact with Pol1A or Pol1B. This region is a part of the primase domain of the protein and slightly overlaps the zinc-finger and RNA polymerase subdomains located within. Additionally, we observed that Arabidopsis Twinkle interacts much more strongly with Pol1A versus Pol1B. 3 Thermophoresis also confirms that the primase domain of Twinkle has higher binding affinity than any other region of the protein. Direct-Coupling-Analysis identified specific residues in Twinkle and the DNA polymerases critical to positive interaction between the two proteins. Conclusions The interaction of Twinkle with Pol1A or Pol1B mimics the minimal DNA replisomes of T7 phage and those present in mammalian mitochondria. However, while T7 and mammals absolutely require their homolog of Twinkle DNA helicase-primase, Arabidopsis Twinkle mutants are seemingly unaffected by this loss. This implies that while Arabidopsis mitochondria mimic minimal replisomes from T7 and mammalian mitochondria, there is an extra level of redundancy specific to loss of Twinkle function.

Keywords

Arabidopsis, Organellar DNA replication, Yeast-Two-Hybrid, Direct-Coupling- Analysis, Thermophoresis

External links

About this resource

• Canonical resource URI: http://purl.org/phylo/treebase/phylows/study/TB2:S24443
• Other versions: Nexus NeXML
• Show BibTeX reference

  		@ARTICLE{TreeBASE2Ref29611,
  	 author = {Stewart  Morley and Antolin  Peralta-Castro and Luis G. Brieba and Justin  Miller and Kai Li  Ong and Perry  Ridge and Amanda  Oliphant and Stephen  Aldous and Brent  Nielsen},
  	 title = {Arabidopsis thaliana organelles mimic the T7 phage DNA replisome with specific interactions between Twinkle protein and DNA polymerases Pol1A and Pol1B},
  	 year = {2019},
  	 keywords = {Arabidopsis, Organellar DNA replication, Yeast-Two-Hybrid, Direct-Coupling- Analysis, Thermophoresis},
  	 doi = {},
  	 url = {http://},
  	 pmid = {},
  	 journal = {BMC Plant Biology},
  	 volume = {},
  	 number = {},
  	 pages = {},
  	 abstract = {Background
  Plant chloroplasts and mitochondria utilize nuclear encoded proteins to replicate their DNA.
  These proteins are purposely built for replication in the organelle environment and are distinct
  from those involved in replication of the nuclear genome. These organelle-localized proteins
  also have ancestral roots in bacterial and bacteriophage genes, supporting the endosymbiotic
  theory of their origin. We examined the interactions between three of these proteins from
  Arabidopsis thaliana: a DNA helicase-primase similar to bacteriophage T7 gp4 protein and
  animal mitochondrial Twinkle, and two DNA polymerases, Pol1A and Pol1B. We used a three-
  pronged approach to analyze the interactions, including Yeast-two-hybrid analysis, Direct
  Coupling Analysis (DCA), and thermophoresis.
  
  Results
  Yeast-two-Hybrid analysis reveals residues 120-295 of Twinkle as the minimal region that can
  still interact with Pol1A or Pol1B. This region is a part of the primase domain of the protein and
  slightly overlaps the zinc-finger and RNA polymerase subdomains located within. Additionally,
  we observed that Arabidopsis Twinkle interacts much more strongly with Pol1A versus Pol1B.
  
  3
  Thermophoresis also confirms that the primase domain of Twinkle has higher binding affinity
  than any other region of the protein. Direct-Coupling-Analysis identified specific residues in
  Twinkle and the DNA polymerases critical to positive interaction between the two proteins.
  
  Conclusions
  The interaction of Twinkle with Pol1A or Pol1B mimics the minimal DNA replisomes of T7 phage
  and those present in mammalian mitochondria. However, while T7 and mammals absolutely
  require their homolog of Twinkle DNA helicase-primase, Arabidopsis Twinkle mutants are
  seemingly unaffected by this loss. This implies that while Arabidopsis mitochondria mimic
  minimal replisomes from T7 and mammalian mitochondria, there is an extra level of
  redundancy specific to loss of Twinkle function.}
  }
  		

• Show RIS reference

  		TY  - JOUR
  ID  - 29611
  AU  - Morley,Stewart 
  AU  - Peralta-Castro,Antolin 
  AU  - Brieba,Luis G.
  AU  - Miller,Justin 
  AU  - Ong,Kai Li 
  AU  - Ridge,Perry 
  AU  - Oliphant,Amanda 
  AU  - Aldous,Stephen 
  AU  - Nielsen,Brent 
  T1  - Arabidopsis thaliana organelles mimic the T7 phage DNA replisome with specific interactions between Twinkle protein and DNA polymerases Pol1A and Pol1B
  PY  - 2019
  KW  - Arabidopsis
  KW  - Organellar DNA replication
  KW  - Yeast-Two-Hybrid
  KW  - Direct-Coupling- Analysis
  KW  - Thermophoresis
  UR  - http://dx.doi.org/
  N2  - Background
  Plant chloroplasts and mitochondria utilize nuclear encoded proteins to replicate their DNA.
  These proteins are purposely built for replication in the organelle environment and are distinct
  from those involved in replication of the nuclear genome. These organelle-localized proteins
  also have ancestral roots in bacterial and bacteriophage genes, supporting the endosymbiotic
  theory of their origin. We examined the interactions between three of these proteins from
  Arabidopsis thaliana: a DNA helicase-primase similar to bacteriophage T7 gp4 protein and
  animal mitochondrial Twinkle, and two DNA polymerases, Pol1A and Pol1B. We used a three-
  pronged approach to analyze the interactions, including Yeast-two-hybrid analysis, Direct
  Coupling Analysis (DCA), and thermophoresis.
  
  Results
  Yeast-two-Hybrid analysis reveals residues 120-295 of Twinkle as the minimal region that can
  still interact with Pol1A or Pol1B. This region is a part of the primase domain of the protein and
  slightly overlaps the zinc-finger and RNA polymerase subdomains located within. Additionally,
  we observed that Arabidopsis Twinkle interacts much more strongly with Pol1A versus Pol1B.
  
  3
  Thermophoresis also confirms that the primase domain of Twinkle has higher binding affinity
  than any other region of the protein. Direct-Coupling-Analysis identified specific residues in
  Twinkle and the DNA polymerases critical to positive interaction between the two proteins.
  
  Conclusions
  The interaction of Twinkle with Pol1A or Pol1B mimics the minimal DNA replisomes of T7 phage
  and those present in mammalian mitochondria. However, while T7 and mammals absolutely
  require their homolog of Twinkle DNA helicase-primase, Arabidopsis Twinkle mutants are
  seemingly unaffected by this loss. This implies that while Arabidopsis mitochondria mimic
  minimal replisomes from T7 and mammalian mitochondria, there is an extra level of
  redundancy specific to loss of Twinkle function.
  L3  - 
  JF  - BMC Plant Biology
  VL  - 
  IS  - 
  ER  -