Quantitative analyses of interactions between SpoVG and RNA/DNA

Timothy C. Saylor; Christina R. Savage; Andrew C. Krusenstjerna; Nerina Jusufovic; Wolfram R. Zückert; Catherine A. Brissette; Md Motaleb; Paula J. Schlax; Brian Stevenson

doi:10.1016/j.bbrc.2023.02.044

Quantitative analyses of interactions between SpoVG and RNA/DNA

Timothy C. Saylor, Christina R. Savage, Andrew C. Krusenstjerna, Nerina Jusufovic, Wolfram R. Zückert, Catherine A. Brissette, Md Motaleb, Paula J. Schlax, Brian Stevenson

Research output: Contribution to journal › Article › peer-review

2 Scopus citations

Abstract

The Borrelia burgdorferi SpoVG protein has previously been found to be a DNA- and RNA-binding protein. To aid in the elucidation of ligand motifs, affinities for numerous RNAs, ssDNAs, and dsDNAs were measured and compared. The loci used in the study were spoVG, glpFKD, erpAB, bb0242, flaB, and ospAB, with particular focus on the untranslated 5′ portion of the mRNAs. Performing binding and competition assays yielded that the 5′ end of spoVG mRNA had the highest affinity while the lowest observed affinity was to the 5’ end of flaB mRNA. Mutagenesis studies of spoVG RNA and ssDNA sequences suggested that the formation of SpoVG-nucleic acid complexes are not entirely dependent on either sequence or structure. Additionally, exchanging uracil for thymine in ssDNAs did not affect protein-nucleic acid complex formation.

Original language	English
Pages (from-to)	40-46
Number of pages	7
Journal	Biochemical and Biophysical Research Communications
Volume	654
DOIs	https://doi.org/10.1016/j.bbrc.2023.02.044
State	Published - Apr 30 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

Funding

These studies were funded by US NIH grant R01 AI144126 . We thank Tatiana Castro-Padovani and Jessamyn Moore for assistance and helpful comments on this manuscript.

Funders	Funder number
National Institutes of Health (NIH)	R01 AI144126
National Institutes of Health (NIH)

Keywords

Borrelia burgdorferi
DNA binding protein
Electrophoretic mobility shift assay (EMSA)
RNA binding protein
SpoVG

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/j.bbrc.2023.02.044

Cite this

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title = "Quantitative analyses of interactions between SpoVG and RNA/DNA",

abstract = "The Borrelia burgdorferi SpoVG protein has previously been found to be a DNA- and RNA-binding protein. To aid in the elucidation of ligand motifs, affinities for numerous RNAs, ssDNAs, and dsDNAs were measured and compared. The loci used in the study were spoVG, glpFKD, erpAB, bb0242, flaB, and ospAB, with particular focus on the untranslated 5′ portion of the mRNAs. Performing binding and competition assays yielded that the 5′ end of spoVG mRNA had the highest affinity while the lowest observed affinity was to the 5{\textquoteright} end of flaB mRNA. Mutagenesis studies of spoVG RNA and ssDNA sequences suggested that the formation of SpoVG-nucleic acid complexes are not entirely dependent on either sequence or structure. Additionally, exchanging uracil for thymine in ssDNAs did not affect protein-nucleic acid complex formation.",

keywords = "Borrelia burgdorferi, DNA binding protein, Electrophoretic mobility shift assay (EMSA), RNA binding protein, SpoVG",

author = "Saylor, {Timothy C.} and Savage, {Christina R.} and Krusenstjerna, {Andrew C.} and Nerina Jusufovic and Z{\"u}ckert, {Wolfram R.} and Brissette, {Catherine A.} and Md Motaleb and Schlax, {Paula J.} and Brian Stevenson",

note = "Publisher Copyright: {\textcopyright} 2023 The Authors",

year = "2023",

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day = "30",

doi = "10.1016/j.bbrc.2023.02.044",

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AU - Saylor, Timothy C.

AU - Savage, Christina R.

AU - Krusenstjerna, Andrew C.

AU - Jusufovic, Nerina

AU - Zückert, Wolfram R.

AU - Brissette, Catherine A.

AU - Motaleb, Md

AU - Schlax, Paula J.

AU - Stevenson, Brian

PY - 2023/4/30

Y1 - 2023/4/30

N2 - The Borrelia burgdorferi SpoVG protein has previously been found to be a DNA- and RNA-binding protein. To aid in the elucidation of ligand motifs, affinities for numerous RNAs, ssDNAs, and dsDNAs were measured and compared. The loci used in the study were spoVG, glpFKD, erpAB, bb0242, flaB, and ospAB, with particular focus on the untranslated 5′ portion of the mRNAs. Performing binding and competition assays yielded that the 5′ end of spoVG mRNA had the highest affinity while the lowest observed affinity was to the 5’ end of flaB mRNA. Mutagenesis studies of spoVG RNA and ssDNA sequences suggested that the formation of SpoVG-nucleic acid complexes are not entirely dependent on either sequence or structure. Additionally, exchanging uracil for thymine in ssDNAs did not affect protein-nucleic acid complex formation.

AB - The Borrelia burgdorferi SpoVG protein has previously been found to be a DNA- and RNA-binding protein. To aid in the elucidation of ligand motifs, affinities for numerous RNAs, ssDNAs, and dsDNAs were measured and compared. The loci used in the study were spoVG, glpFKD, erpAB, bb0242, flaB, and ospAB, with particular focus on the untranslated 5′ portion of the mRNAs. Performing binding and competition assays yielded that the 5′ end of spoVG mRNA had the highest affinity while the lowest observed affinity was to the 5’ end of flaB mRNA. Mutagenesis studies of spoVG RNA and ssDNA sequences suggested that the formation of SpoVG-nucleic acid complexes are not entirely dependent on either sequence or structure. Additionally, exchanging uracil for thymine in ssDNAs did not affect protein-nucleic acid complex formation.

KW - Borrelia burgdorferi

KW - DNA binding protein

KW - Electrophoretic mobility shift assay (EMSA)

KW - RNA binding protein

KW - SpoVG

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JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

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Quantitative analyses of interactions between SpoVG and RNA/DNA

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

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