Developmental and tissue specific changes of ubiquitin forms in drosophila melanogaster

Ágota Nagy, Levente Kovács, Zoltán Lipinszki, Margit Pál, P. Deák

Research output: Contribution to journalArticle

Abstract

In most Eukaryotes, ubiquitin either exists as free monoubiquitin or as a molecule that is covalently linked to other proteins. These two forms cycle between each other and due to the concerted antagonistic activity of ubiquitylating and deubiquitylating enzymes, an intracellular ubiquitin equilibrium is maintained that is essential for normal biological function. However, measuring the level and ratio of these forms of ubiquitin has been difficult and time consuming. In this paper, we have adapted a simple immunoblotting technique to monitor ubiquitin content and equilibrium dynamics in different developmental stages and tissues of Drosophila. Our data show that the level of total ubiquitin is distinct in different developmental stages, lowest at the larval-pupal transition and in three days old adult males, and highest in first instar larvae. Interestingly, the ratio of free mono-ubiquitin remains within 30–50% range of the total throughout larval development, but peaks to 70–80% at the larval-pupal and the pupal-adult transitions. It stays within the 70–80% range in adults. In developmentally and physiologically active tissues, the ratio of free ubiquitin is similarly high, most likely reflecting a high demand for ubiquitin availability. We also used this method to demonstrate the disruption of the finely tuned ubiquitin equilibrium by the abolition of proteasome function or the housekeeping deubiquitylase, Usp5. Our data support the notion that the ubiquitin equilibrium is regulated by tissue- and developmental stage-specific mechanisms.

Original languageEnglish
Article numbere0209080
JournalPLoS One
Volume13
Issue number12
DOIs
Publication statusPublished - Dec 1 2018

Fingerprint

ubiquitin
Ubiquitin
Drosophila melanogaster
Tissue
developmental stages
tissues
Housekeeping
proteasome endopeptidase complex
Proteasome Endopeptidase Complex
immunoblotting
Eukaryota
Immunoblotting
larval development
Drosophila
Larva
eukaryotic cells
instars
Availability
Molecules
monitoring

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Developmental and tissue specific changes of ubiquitin forms in drosophila melanogaster. / Nagy, Ágota; Kovács, Levente; Lipinszki, Zoltán; Pál, Margit; Deák, P.

In: PLoS One, Vol. 13, No. 12, e0209080, 01.12.2018.

Research output: Contribution to journalArticle

Nagy, Ágota ; Kovács, Levente ; Lipinszki, Zoltán ; Pál, Margit ; Deák, P. / Developmental and tissue specific changes of ubiquitin forms in drosophila melanogaster. In: PLoS One. 2018 ; Vol. 13, No. 12.
@article{d21de640f36d40f2987f2ff8ae5065c2,
title = "Developmental and tissue specific changes of ubiquitin forms in drosophila melanogaster",
abstract = "In most Eukaryotes, ubiquitin either exists as free monoubiquitin or as a molecule that is covalently linked to other proteins. These two forms cycle between each other and due to the concerted antagonistic activity of ubiquitylating and deubiquitylating enzymes, an intracellular ubiquitin equilibrium is maintained that is essential for normal biological function. However, measuring the level and ratio of these forms of ubiquitin has been difficult and time consuming. In this paper, we have adapted a simple immunoblotting technique to monitor ubiquitin content and equilibrium dynamics in different developmental stages and tissues of Drosophila. Our data show that the level of total ubiquitin is distinct in different developmental stages, lowest at the larval-pupal transition and in three days old adult males, and highest in first instar larvae. Interestingly, the ratio of free mono-ubiquitin remains within 30–50{\%} range of the total throughout larval development, but peaks to 70–80{\%} at the larval-pupal and the pupal-adult transitions. It stays within the 70–80{\%} range in adults. In developmentally and physiologically active tissues, the ratio of free ubiquitin is similarly high, most likely reflecting a high demand for ubiquitin availability. We also used this method to demonstrate the disruption of the finely tuned ubiquitin equilibrium by the abolition of proteasome function or the housekeeping deubiquitylase, Usp5. Our data support the notion that the ubiquitin equilibrium is regulated by tissue- and developmental stage-specific mechanisms.",
author = "{\'A}gota Nagy and Levente Kov{\'a}cs and Zolt{\'a}n Lipinszki and Margit P{\'a}l and P. De{\'a}k",
year = "2018",
month = "12",
day = "1",
doi = "10.1371/journal.pone.0209080",
language = "English",
volume = "13",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "12",

}

TY - JOUR

T1 - Developmental and tissue specific changes of ubiquitin forms in drosophila melanogaster

AU - Nagy, Ágota

AU - Kovács, Levente

AU - Lipinszki, Zoltán

AU - Pál, Margit

AU - Deák, P.

PY - 2018/12/1

Y1 - 2018/12/1

N2 - In most Eukaryotes, ubiquitin either exists as free monoubiquitin or as a molecule that is covalently linked to other proteins. These two forms cycle between each other and due to the concerted antagonistic activity of ubiquitylating and deubiquitylating enzymes, an intracellular ubiquitin equilibrium is maintained that is essential for normal biological function. However, measuring the level and ratio of these forms of ubiquitin has been difficult and time consuming. In this paper, we have adapted a simple immunoblotting technique to monitor ubiquitin content and equilibrium dynamics in different developmental stages and tissues of Drosophila. Our data show that the level of total ubiquitin is distinct in different developmental stages, lowest at the larval-pupal transition and in three days old adult males, and highest in first instar larvae. Interestingly, the ratio of free mono-ubiquitin remains within 30–50% range of the total throughout larval development, but peaks to 70–80% at the larval-pupal and the pupal-adult transitions. It stays within the 70–80% range in adults. In developmentally and physiologically active tissues, the ratio of free ubiquitin is similarly high, most likely reflecting a high demand for ubiquitin availability. We also used this method to demonstrate the disruption of the finely tuned ubiquitin equilibrium by the abolition of proteasome function or the housekeeping deubiquitylase, Usp5. Our data support the notion that the ubiquitin equilibrium is regulated by tissue- and developmental stage-specific mechanisms.

AB - In most Eukaryotes, ubiquitin either exists as free monoubiquitin or as a molecule that is covalently linked to other proteins. These two forms cycle between each other and due to the concerted antagonistic activity of ubiquitylating and deubiquitylating enzymes, an intracellular ubiquitin equilibrium is maintained that is essential for normal biological function. However, measuring the level and ratio of these forms of ubiquitin has been difficult and time consuming. In this paper, we have adapted a simple immunoblotting technique to monitor ubiquitin content and equilibrium dynamics in different developmental stages and tissues of Drosophila. Our data show that the level of total ubiquitin is distinct in different developmental stages, lowest at the larval-pupal transition and in three days old adult males, and highest in first instar larvae. Interestingly, the ratio of free mono-ubiquitin remains within 30–50% range of the total throughout larval development, but peaks to 70–80% at the larval-pupal and the pupal-adult transitions. It stays within the 70–80% range in adults. In developmentally and physiologically active tissues, the ratio of free ubiquitin is similarly high, most likely reflecting a high demand for ubiquitin availability. We also used this method to demonstrate the disruption of the finely tuned ubiquitin equilibrium by the abolition of proteasome function or the housekeeping deubiquitylase, Usp5. Our data support the notion that the ubiquitin equilibrium is regulated by tissue- and developmental stage-specific mechanisms.

UR - http://www.scopus.com/inward/record.url?scp=85058487442&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058487442&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0209080

DO - 10.1371/journal.pone.0209080

M3 - Article

VL - 13

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 12

M1 - e0209080

ER -