The repeated 36 amino acid motif of Chlamydia trachomatis Hc2 protein binds to the major groove of DNA

Odete Sofia Lopes Gonçalves, Gunna Christiansen, Arne Holm, Bjørn Herrmann, Markus Klintstedt, Steffen B Petersen, Svend Birkelund

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

The gram-negative, obligate intracellular human pathogen, Chlamydia trachomatis has a bi-phasic developmental cycle. The histone H1-like C. trachomatis DNA binding protein, Hc2, is produced late during the developmental cycle when the dividing reticulate body transforms into the smaller, metabolically inactive elementary body. Together with Hc1, the two proteins compact the chlamydial chromosome and arrest replication and transcription. Hc2 is heterogeneous in length due to variation in the number of lysine rich pentamers. Six pentamers and one hexamer constitute a 36 amino acid long repetitive unit that, in spite of variations, is unique for Chlamydiaceae. Using synthetic peptides, the DNA-binding capacity of the 36 amino acid peptide and that of a randomized peptide was analyzed. Both peptides bound and compacted plasmid DNA, however, electron microscopy of peptide/DNA complexes showed major differences in the resulting aggregated structures. Fluorescence spectroscopy was used to analyze the binding. After complexing plasmid DNA with each of three different intercalating dyes, increasing amounts of peptides were added and fluorescence spectroscopy performed. The major groove binder, methyl green, was displaced by both peptides at low concentrations, while the minor groove binder, Hoechts, and the intercalating dye, Ethidium Bromide, were displaced only at high concentrations of peptides.

OriginalsprogEngelsk
TidsskriftResearch in Microbiology
Vol/bind170
Udgave nummer6-7
Sider (fra-til)256-262
Antal sider7
ISSN0923-2508
DOI
StatusUdgivet - 1 sep. 2019

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Amino Acid Motifs
Peptides
DNA
Chlamydia trachomatis
Fluorescence Spectrometry
Chlamydiaceae
Plasmids
Coloring Agents
Methyl Green
Amino Acids
Ethidium
Chlamydia trachomatis HctB protein
DNA-Binding Proteins
Histones
Lysine
Electron Microscopy
Chromosomes

Citer dette

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title = "The repeated 36 amino acid motif of Chlamydia trachomatis Hc2 protein binds to the major groove of DNA",
abstract = "The gram-negative, obligate intracellular human pathogen, Chlamydia trachomatis has a bi-phasic developmental cycle. The histone H1-like C. trachomatis DNA binding protein, Hc2, is produced late during the developmental cycle when the dividing reticulate body transforms into the smaller, metabolically inactive elementary body. Together with Hc1, the two proteins compact the chlamydial chromosome and arrest replication and transcription. Hc2 is heterogeneous in length due to variation in the number of lysine rich pentamers. Six pentamers and one hexamer constitute a 36 amino acid long repetitive unit that, in spite of variations, is unique for Chlamydiaceae. Using synthetic peptides, the DNA-binding capacity of the 36 amino acid peptide and that of a randomized peptide was analyzed. Both peptides bound and compacted plasmid DNA, however, electron microscopy of peptide/DNA complexes showed major differences in the resulting aggregated structures. Fluorescence spectroscopy was used to analyze the binding. After complexing plasmid DNA with each of three different intercalating dyes, increasing amounts of peptides were added and fluorescence spectroscopy performed. The major groove binder, methyl green, was displaced by both peptides at low concentrations, while the minor groove binder, Hoechts, and the intercalating dye, Ethidium Bromide, were displaced only at high concentrations of peptides.",
keywords = "Chlamydia trachomatis, DNA packing, Fluorescence spectroscopy, Hc2, Histone H1-like protein, Methyl green",
author = "{Lopes Gon{\cc}alves}, {Odete Sofia} and Gunna Christiansen and Arne Holm and Bj{\o}rn Herrmann and Markus Klintstedt and Petersen, {Steffen B} and Svend Birkelund",
note = "Copyright {\circledC} 2019 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.",
year = "2019",
month = "9",
day = "1",
doi = "10.1016/j.resmic.2019.08.002",
language = "English",
volume = "170",
pages = "256--262",
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The repeated 36 amino acid motif of Chlamydia trachomatis Hc2 protein binds to the major groove of DNA. / Lopes Gonçalves, Odete Sofia; Christiansen, Gunna; Holm, Arne; Herrmann, Bjørn; Klintstedt, Markus; Petersen, Steffen B; Birkelund, Svend.

I: Research in Microbiology, Bind 170, Nr. 6-7, 01.09.2019, s. 256-262.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - The repeated 36 amino acid motif of Chlamydia trachomatis Hc2 protein binds to the major groove of DNA

AU - Lopes Gonçalves, Odete Sofia

AU - Christiansen, Gunna

AU - Holm, Arne

AU - Herrmann, Bjørn

AU - Klintstedt, Markus

AU - Petersen, Steffen B

AU - Birkelund, Svend

N1 - Copyright © 2019 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

PY - 2019/9/1

Y1 - 2019/9/1

N2 - The gram-negative, obligate intracellular human pathogen, Chlamydia trachomatis has a bi-phasic developmental cycle. The histone H1-like C. trachomatis DNA binding protein, Hc2, is produced late during the developmental cycle when the dividing reticulate body transforms into the smaller, metabolically inactive elementary body. Together with Hc1, the two proteins compact the chlamydial chromosome and arrest replication and transcription. Hc2 is heterogeneous in length due to variation in the number of lysine rich pentamers. Six pentamers and one hexamer constitute a 36 amino acid long repetitive unit that, in spite of variations, is unique for Chlamydiaceae. Using synthetic peptides, the DNA-binding capacity of the 36 amino acid peptide and that of a randomized peptide was analyzed. Both peptides bound and compacted plasmid DNA, however, electron microscopy of peptide/DNA complexes showed major differences in the resulting aggregated structures. Fluorescence spectroscopy was used to analyze the binding. After complexing plasmid DNA with each of three different intercalating dyes, increasing amounts of peptides were added and fluorescence spectroscopy performed. The major groove binder, methyl green, was displaced by both peptides at low concentrations, while the minor groove binder, Hoechts, and the intercalating dye, Ethidium Bromide, were displaced only at high concentrations of peptides.

AB - The gram-negative, obligate intracellular human pathogen, Chlamydia trachomatis has a bi-phasic developmental cycle. The histone H1-like C. trachomatis DNA binding protein, Hc2, is produced late during the developmental cycle when the dividing reticulate body transforms into the smaller, metabolically inactive elementary body. Together with Hc1, the two proteins compact the chlamydial chromosome and arrest replication and transcription. Hc2 is heterogeneous in length due to variation in the number of lysine rich pentamers. Six pentamers and one hexamer constitute a 36 amino acid long repetitive unit that, in spite of variations, is unique for Chlamydiaceae. Using synthetic peptides, the DNA-binding capacity of the 36 amino acid peptide and that of a randomized peptide was analyzed. Both peptides bound and compacted plasmid DNA, however, electron microscopy of peptide/DNA complexes showed major differences in the resulting aggregated structures. Fluorescence spectroscopy was used to analyze the binding. After complexing plasmid DNA with each of three different intercalating dyes, increasing amounts of peptides were added and fluorescence spectroscopy performed. The major groove binder, methyl green, was displaced by both peptides at low concentrations, while the minor groove binder, Hoechts, and the intercalating dye, Ethidium Bromide, were displaced only at high concentrations of peptides.

KW - Chlamydia trachomatis

KW - DNA packing

KW - Fluorescence spectroscopy

KW - Hc2

KW - Histone H1-like protein

KW - Methyl green

UR - http://www.scopus.com/inward/record.url?scp=85071246981&partnerID=8YFLogxK

U2 - 10.1016/j.resmic.2019.08.002

DO - 10.1016/j.resmic.2019.08.002

M3 - Journal article

VL - 170

SP - 256

EP - 262

JO - Research in Microbiology

JF - Research in Microbiology

SN - 0923-2508

IS - 6-7

ER -