Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase

Marek Wojciechowski, Dominik Rafalski, Robert Kucharski, Katarzyna Misztal, Joanna Maleszka, Matthias Bochtler, Ryszard Maleszka

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

In mammals, a family of TET enzymes producing oxidized forms of 5-methylcyto-sine (5mC) plays an important role in modulating DNA demethylation dynamics. In contrast, nothingisknown about the functionofasingle TETorthologue present in invertebrates. Here, we show that the honeybee TET (AmTET) catalytic domain has dioxygenase activity and converts 5mC to 5-hydroxymethylcytosine (5hmC) in a HEK293T cell assay. In vivo, the levels of 5hmC are condition-dependent and relatively low, but in testes and ovaries 5hmC is present at approximately 7-10% of the total level of 5mC, which is comparable to that reported for certain mammalian cells types. AmTET is alternatively spliced and highly expressed throughout development and in adult tissues with the highest expression found in adult brains. Our findings reveal an additional level of flexible genomic modifications in the honeybee that may be important for the selection of multiple pathways controlling contrasting phenotypic outcomes in this species. In a broader context, our study extends the current, mammalian-centred attention to TET-driven DNA hydroxymethylation to an easily manageable organism with attractive and unique biology.

Original languageEnglish
Article number140110
Pages (from-to)1-9
Number of pages9
JournalOpen Biology
Volume4
Issue number8
DOIs
Publication statusPublished - 6 Aug 2014
Externally publishedYes

Cite this

Wojciechowski, M., Rafalski, D., Kucharski, R., Misztal, K., Maleszka, J., Bochtler, M., & Maleszka, R. (2014). Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase. Open Biology, 4(8), 1-9. [140110]. https://doi.org/10.1098/rsob.140110
Wojciechowski, Marek ; Rafalski, Dominik ; Kucharski, Robert ; Misztal, Katarzyna ; Maleszka, Joanna ; Bochtler, Matthias ; Maleszka, Ryszard. / Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase. In: Open Biology. 2014 ; Vol. 4, No. 8. pp. 1-9.
@article{88e23e33fbca461ea2106a4603c269e6,
title = "Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase",
abstract = "In mammals, a family of TET enzymes producing oxidized forms of 5-methylcyto-sine (5mC) plays an important role in modulating DNA demethylation dynamics. In contrast, nothingisknown about the functionofasingle TETorthologue present in invertebrates. Here, we show that the honeybee TET (AmTET) catalytic domain has dioxygenase activity and converts 5mC to 5-hydroxymethylcytosine (5hmC) in a HEK293T cell assay. In vivo, the levels of 5hmC are condition-dependent and relatively low, but in testes and ovaries 5hmC is present at approximately 7-10{\%} of the total level of 5mC, which is comparable to that reported for certain mammalian cells types. AmTET is alternatively spliced and highly expressed throughout development and in adult tissues with the highest expression found in adult brains. Our findings reveal an additional level of flexible genomic modifications in the honeybee that may be important for the selection of multiple pathways controlling contrasting phenotypic outcomes in this species. In a broader context, our study extends the current, mammalian-centred attention to TET-driven DNA hydroxymethylation to an easily manageable organism with attractive and unique biology.",
keywords = "Brain plasticity, Demethylation, Epigenetic code, Epigenomics, Phenotypic polymorphism, Social insect, Ovary/enzymology, Alternative Splicing, Epigenesis, Genetic, Humans, Cytosine/analogs & derivatives, Molecular Sequence Data, Male, Insect Proteins/genetics, Bees, DNA Methylation, Gene Expression Regulation, Developmental, HEK293 Cells, Female, Transgenes, Amino Acid Sequence, Testis/enzymology, Catalytic Domain, Brain/enzymology, Dioxygenases/genetics, 5-Methylcytosine/metabolism, Organ Specificity, Sequence Alignment, Animals",
author = "Marek Wojciechowski and Dominik Rafalski and Robert Kucharski and Katarzyna Misztal and Joanna Maleszka and Matthias Bochtler and Ryszard Maleszka",
year = "2014",
month = "8",
day = "6",
doi = "10.1098/rsob.140110",
language = "English",
volume = "4",
pages = "1--9",
journal = "Open Biology",
issn = "2046-2441",
publisher = "Royal Society Publishing",
number = "8",

}

Wojciechowski, M, Rafalski, D, Kucharski, R, Misztal, K, Maleszka, J, Bochtler, M & Maleszka, R 2014, 'Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase', Open Biology, vol. 4, no. 8, 140110, pp. 1-9. https://doi.org/10.1098/rsob.140110

Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase. / Wojciechowski, Marek; Rafalski, Dominik; Kucharski, Robert; Misztal, Katarzyna; Maleszka, Joanna; Bochtler, Matthias; Maleszka, Ryszard.

In: Open Biology, Vol. 4, No. 8, 140110, 06.08.2014, p. 1-9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase

AU - Wojciechowski, Marek

AU - Rafalski, Dominik

AU - Kucharski, Robert

AU - Misztal, Katarzyna

AU - Maleszka, Joanna

AU - Bochtler, Matthias

AU - Maleszka, Ryszard

PY - 2014/8/6

Y1 - 2014/8/6

N2 - In mammals, a family of TET enzymes producing oxidized forms of 5-methylcyto-sine (5mC) plays an important role in modulating DNA demethylation dynamics. In contrast, nothingisknown about the functionofasingle TETorthologue present in invertebrates. Here, we show that the honeybee TET (AmTET) catalytic domain has dioxygenase activity and converts 5mC to 5-hydroxymethylcytosine (5hmC) in a HEK293T cell assay. In vivo, the levels of 5hmC are condition-dependent and relatively low, but in testes and ovaries 5hmC is present at approximately 7-10% of the total level of 5mC, which is comparable to that reported for certain mammalian cells types. AmTET is alternatively spliced and highly expressed throughout development and in adult tissues with the highest expression found in adult brains. Our findings reveal an additional level of flexible genomic modifications in the honeybee that may be important for the selection of multiple pathways controlling contrasting phenotypic outcomes in this species. In a broader context, our study extends the current, mammalian-centred attention to TET-driven DNA hydroxymethylation to an easily manageable organism with attractive and unique biology.

AB - In mammals, a family of TET enzymes producing oxidized forms of 5-methylcyto-sine (5mC) plays an important role in modulating DNA demethylation dynamics. In contrast, nothingisknown about the functionofasingle TETorthologue present in invertebrates. Here, we show that the honeybee TET (AmTET) catalytic domain has dioxygenase activity and converts 5mC to 5-hydroxymethylcytosine (5hmC) in a HEK293T cell assay. In vivo, the levels of 5hmC are condition-dependent and relatively low, but in testes and ovaries 5hmC is present at approximately 7-10% of the total level of 5mC, which is comparable to that reported for certain mammalian cells types. AmTET is alternatively spliced and highly expressed throughout development and in adult tissues with the highest expression found in adult brains. Our findings reveal an additional level of flexible genomic modifications in the honeybee that may be important for the selection of multiple pathways controlling contrasting phenotypic outcomes in this species. In a broader context, our study extends the current, mammalian-centred attention to TET-driven DNA hydroxymethylation to an easily manageable organism with attractive and unique biology.

KW - Brain plasticity

KW - Demethylation

KW - Epigenetic code

KW - Epigenomics

KW - Phenotypic polymorphism

KW - Social insect

KW - Ovary/enzymology

KW - Alternative Splicing

KW - Epigenesis, Genetic

KW - Humans

KW - Cytosine/analogs & derivatives

KW - Molecular Sequence Data

KW - Male

KW - Insect Proteins/genetics

KW - Bees

KW - DNA Methylation

KW - Gene Expression Regulation, Developmental

KW - HEK293 Cells

KW - Female

KW - Transgenes

KW - Amino Acid Sequence

KW - Testis/enzymology

KW - Catalytic Domain

KW - Brain/enzymology

KW - Dioxygenases/genetics

KW - 5-Methylcytosine/metabolism

KW - Organ Specificity

KW - Sequence Alignment

KW - Animals

UR - http://www.scopus.com/inward/record.url?scp=84923566739&partnerID=8YFLogxK

UR - http://www.mendeley.com/research/insights-dna-hydroxymethylation-honeybee-indepth-analyses-tet-dioxygenase-1

U2 - 10.1098/rsob.140110

DO - 10.1098/rsob.140110

M3 - Article

VL - 4

SP - 1

EP - 9

JO - Open Biology

JF - Open Biology

SN - 2046-2441

IS - 8

M1 - 140110

ER -

Wojciechowski M, Rafalski D, Kucharski R, Misztal K, Maleszka J, Bochtler M et al. Insights into DNA hydroxymethylation in the honeybee from in-depth analyses of TET dioxygenase. Open Biology. 2014 Aug 6;4(8):1-9. 140110. https://doi.org/10.1098/rsob.140110