Fly versus man: evolutionary impairment of nucleolar targeting affects the degradome of Drosophila's Taspase1

Autoren

Désirée Wünsch
Angelina Hahlbrock
Christina Heiselmayer
Sandra Bäcker
Patrick Heun
Dorothee Goesswein
Walter Stöcker
Tanja Schirmeister
Günter Schneider
Oliver H Krämer
Shirley K Knauer
Roland H Stauber

DOI

10.1096/fj.14-262451

eISSN

1530-6860

ISSN

0892-6638

Ausgabe der Veröffentlichung

5

Zeitschrift

The FASEB Journal

Sprache

en

Online publication date

2015

Paginierung

1973 - 1985

Datum der Veröffentlichung

2015

Status

Published

Herausgeber

Wiley

Herausgeber URL

http://dx.doi.org/10.1096/fj.14-262451

Datum der Datenerfassung

2022

Titel

Fly<i>versus</i>man: evolutionary impairment of nucleolar targeting affects the degradome of<i>Drosophila's</i>Taspase1

Ausgabe der Zeitschrift

29

Datenquelle: Crossref

Abstract

Human Taspase1 is essential for development and cancer by processing critical regulators, such as the mixed-lineage leukemia protein. Likewise, its ortholog, trithorax, is cleaved by Drosophila Taspase1 (dTaspase1), implementing a functional coevolution. To uncover novel mechanism regulating protease function, we performed a functional analysis of dTaspase1 and its comparison to the human ortholog. dTaspase1 contains an essential nucleophile threonine(195), catalyzing cis cleavage into its α- and β-subunits. A cell-based assay combined with alanine scanning mutagenesis demonstrated that the target cleavage motif for dTaspase1 (Q(3)[F/I/L/M](2)D(1)↓G(1')X(2')X(3')) differs significantly from the human ortholog (Q(3)[F,I,L,V](2)D(1)↓G(1')x(2')D(3')D(4')), predicting an enlarged degradome containing 70 substrates for Drosophila. In contrast to human Taspase1, dTaspase1 shows no discrete localization to the nucleus/nucleolus due to the lack of the importin-α/nucleophosmin1 interaction domain (NoLS) conserved in all vertebrates. Consequently, dTaspase1 interacts with neither the Drosophila nucleoplasmin-like protein nor human nucleophosmin1. The impact of localization on the protease's degradome was confirmed by demonstrating that dTaspase1 did not efficiently process nuclear substrates, such as upstream stimulatory factor 2. However, genetic introduction of the NoLS into dTaspase1 restored its nucleolar localization, nucleophosmin1 interaction, and efficient cleavage of nuclear substrates. We report that evolutionary functional divergence separating vertebrates from invertebrates can be achieved for proteases by a transport/localization-regulated mechanism.

Addresses

*Molecular and Cellular Oncology, Ear, Nose and Throat Department, University Medical Center of Mainz, Mainz, Germany; Institute for Molecular Biology, Centre for Medical Biotechnology, Mainz Scientific Screening Center UG & Co. KG, University of Duisburg-Essen, Essen, Germany; Max Planck Institute of Immunobiology and Epigenetics, Freiburg, Germany; Cell and Matrix Biology, Institute for Zoology, and Institute for Pharmacy and Biochemistry, University of Mainz, Mainz, Germany; II. Medizinische Klinik, Technische Universität München, Munich, Germany; and Department of Toxicology, University Medical Center, Mainz, Germany wuensch@uni-mainz.de rstauber@uni-mainz.de.

Autoren

Désirée Wünsch
Angelina Hahlbrock
Christina Heiselmayer
Sandra Bäcker
Patrick Heun
Dorothee Goesswein
Walter Stöcker
Tanja Schirmeister
Günter Schneider
Oliver H Krämer
Shirley K Knauer
Roland H Stauber

DOI

10.1096/fj.14-262451

eISSN

1530-6860

Externe Identifier

PubMed Identifier: 25634959

Funding acknowledgements

Fritz Thyssen Stiftung:

Open access

false

ISSN

0892-6638

Ausgabe der Veröffentlichung

5

Zeitschrift

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

Schlüsselwörter

Cells, Cultured
Cell Nucleus
Cell Nucleolus
Animals
Humans
Drosophila
Peptide Hydrolases
Endopeptidases
Drosophila Proteins
Microscopy, Confocal
Fluorescent Antibody Technique
Blotting, Western
Mutagenesis, Site-Directed
Immunoprecipitation
Phylogeny
Signal Transduction
Amino Acid Sequence
Sequence Homology, Amino Acid
Protein Transport
Molecular Sequence Data
Male
Biological Evolution
Proteolysis

Sprache

eng

Medium

Print-Electronic

Online publication date

2015

Paginierung

1973 - 1985

Datum der Veröffentlichung

2015

Status

Published

Datum der Datenerfassung

2015

Titel

Fly versus man: evolutionary impairment of nucleolar targeting affects the degradome of Drosophila's Taspase1.

Sub types

Comparative Study
Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

29

Datenquelle: Europe PubMed Central

Abstract

Human Taspase1 is essential for development and cancer by processing critical regulators, such as the mixed-lineage leukemia protein. Likewise, its ortholog, trithorax, is cleaved by Drosophila Taspase1 (dTaspase1), implementing a functional coevolution. To uncover novel mechanism regulating protease function, we performed a functional analysis of dTaspase1 and its comparison to the human ortholog. dTaspase1 contains an essential nucleophile threonine(195), catalyzing cis cleavage into its α- and β-subunits. A cell-based assay combined with alanine scanning mutagenesis demonstrated that the target cleavage motif for dTaspase1 (Q(3)[F/I/L/M](2)D(1)↓G(1')X(2')X(3')) differs significantly from the human ortholog (Q(3)[F,I,L,V](2)D(1)↓G(1')x(2')D(3')D(4')), predicting an enlarged degradome containing 70 substrates for Drosophila. In contrast to human Taspase1, dTaspase1 shows no discrete localization to the nucleus/nucleolus due to the lack of the importin-α/nucleophosmin1 interaction domain (NoLS) conserved in all vertebrates. Consequently, dTaspase1 interacts with neither the Drosophila nucleoplasmin-like protein nor human nucleophosmin1. The impact of localization on the protease's degradome was confirmed by demonstrating that dTaspase1 did not efficiently process nuclear substrates, such as upstream stimulatory factor 2. However, genetic introduction of the NoLS into dTaspase1 restored its nucleolar localization, nucleophosmin1 interaction, and efficient cleavage of nuclear substrates. We report that evolutionary functional divergence separating vertebrates from invertebrates can be achieved for proteases by a transport/localization-regulated mechanism.

Date of acceptance

2014

Autoren

Désirée Wünsch
Angelina Hahlbrock
Christina Heiselmayer
Sandra Bäcker
Patrick Heun
Dorothee Goesswein
Walter Stöcker
Tanja Schirmeister
Günter Schneider
Oliver H Krämer
Shirley K Knauer
Roland H Stauber

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/25634959

DOI

10.1096/fj.14-262451

eISSN

1530-6860

Ausgabe der Veröffentlichung

5

Zeitschrift

FASEB J

Schlüsselwörter

cancer
development
leukemia
protease
threonine aspartase
Amino Acid Sequence
Animals
Biological Evolution
Blotting, Western
Cell Nucleolus
Cell Nucleus
Cells, Cultured
Drosophila
Drosophila Proteins
Endopeptidases
Fluorescent Antibody Technique
Humans
Immunoprecipitation
Male
Microscopy, Confocal
Molecular Sequence Data
Mutagenesis, Site-Directed
Peptide Hydrolases
Phylogeny
Protein Transport
Proteolysis
Sequence Homology, Amino Acid
Signal Transduction

Sprache

eng

Country

United States

Paginierung

1973 - 1985

PII

fj.14-262451

Datum der Veröffentlichung

2015

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2015

Titel

Fly versus man: evolutionary impairment of nucleolar targeting affects the degradome of Drosophila's Taspase1.

Sub types

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

29

Datenquelle: PubMed

Fly versus man: evolutionary impairment of nucleolar targeting affects the degradome of Drosophila's Taspase1

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