scholarly journals Evolution of an Amniote-Specific Mechanism for Modulating Ubiquitin Signaling via Phosphoregulation of the E2 Enzyme UBE2D3

2020 ◽  
Vol 37 (7) ◽  
pp. 1986-2001
Author(s):  
Monica Roman-Trufero ◽  
Constance M Ito ◽  
Conrado Pedebos ◽  
Indiana Magdalou ◽  
Yi-Fang Wang ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Phosphorylation Site ◽  
Embryonic Stem ◽  
Primitive Endoderm ◽  
E3 Ligases ◽  
Proximity Ligation ◽  
Endoderm Differentiation ◽  
Protein Sequence Comparison ◽  
Helical Region

Abstract Genetic variation in the enzymes that catalyze posttranslational modification of proteins is a potentially important source of phenotypic variation during evolution. Ubiquitination is one such modification that affects turnover of virtually all of the proteins in the cell in addition to roles in signaling and epigenetic regulation. UBE2D3 is a promiscuous E2 enzyme, which acts as an ubiquitin donor for E3 ligases that catalyze ubiquitination of developmentally important proteins. We have used protein sequence comparison of UBE2D3 orthologs to identify a position in the C-terminal α-helical region of UBE2D3 that is occupied by a conserved serine in amniotes and by alanine in anamniote vertebrate and invertebrate lineages. Acquisition of the serine (S138) in the common ancestor to modern amniotes created a phosphorylation site for Aurora B. Phosphorylation of S138 disrupts the structure of UBE2D3 and reduces the level of the protein in mouse embryonic stem cells (ESCs). Substitution of S138 with the anamniote alanine (S138A) increases the level of UBE2D3 in ESCs as well as being a gain of function early embryonic lethal mutation in mice. When mutant S138A ESCs were differentiated into extraembryonic primitive endoderm, levels of the PDGFRα and FGFR1 receptor tyrosine kinases were reduced and primitive endoderm differentiation was compromised. Proximity ligation analysis showed increased interaction between UBE2D3 and the E3 ligase CBL and between CBL and the receptor tyrosine kinases. Our results identify a sequence change that altered the ubiquitination landscape at the base of the amniote lineage with potential effects on amniote biology and evolution.

scholarly journals Evolution of an amniote-specific mechanism for modulating ubiquitin signalling via phosphoregulation of the E2 enzyme UBE2D3

2019 ◽  
Author(s):  
Monica Roman-Trufero ◽  
Constance M Ito ◽  
Conrado Pedebos ◽  
Indiana Magdalou ◽  
Yi-Fang Wang ◽  
...  
Keyword(s):  
Tyrosine Kinases ◽  
Es Cells ◽  
Phosphorylation Site ◽  
Post Translational Modification ◽  
E3 Ligases ◽  
Mouse Es Cells ◽  
Proximity Ligation ◽  
The Common ◽  
Protein Sequence Comparison ◽  
Helical Region

AbstractGenetic variation in the enzymes that catalyse post-translational modification of proteins is a potentially important source of phenotypic variation during evolution. Ubiquitination is one such modification that affects turnover of virtually all of the proteins in the cell in addition to roles in signalling and epigenetic regulation. UBE2D3 is a promiscuous E2 enzyme that acts as a ubiquitin donor for E3 ligases that catalyse ubiquitination of developmentally important proteins. We have used protein sequence comparison of UBE2D3 orthologues to identify a position in the C-terminal α-helical region of UBE2D3 that is occupied by a conserved serine in amniotes and by alanine in anamniote vertebrate and invertebrate lineages. Acquisition of the serine (S138) in the common ancestor to modern amniotes created a phosphorylation site for Aurora B. Phosphorylation of S138 disrupts the structure of UBE2D3 and reduces the level of the protein in mouse ES cells (ESCs). Substitution of S138 with the anamniote alanine (S138A) increases the level of UBE2D3 in ESCs as well as being a gain of function early embryonic lethal in mice. When mutant S138A ESCs were differentiated into extra-embryonic primitive endoderm (PrE), levels of the PDGFRα and FGFR1 receptor tyrosine kinases (RTKs) were reduced and PreE differentiation was compromised. Proximity ligation analysis showed increased interaction between UBE2D3 and the E3 ligase CBL and between CBL and the RTKs. Our results identify a sequence change that altered the ubiquitination landscape at the base of the amniote lineage with potential effects on amniote biology and evolution.

scholarly journals The Grb2/Mek Pathway Represses Nanog in Murine Embryonic Stem Cells

2006 ◽  
Vol 26 (20) ◽  
pp. 7539-7549 ◽  
Author(s):  
Takashi Hamazaki ◽  
Sarah M. Kehoe ◽  
Toru Nakano ◽  
Naohiro Terada
Keyword(s):  
Es Cells ◽  
Tyrosine Phosphatase ◽  
Embryonic Stem ◽  
Homeobox Gene ◽  
Cell Aggregation ◽  
Cell Aggregates ◽  
Es Cell ◽  
Sodium Vanadate ◽  
Primitive Endoderm ◽  
Endoderm Differentiation

ABSTRACT The homeobox gene Nanog is a key intrinsic determinant of self renewal in embryonic stem (ES) cells, and its repression leads ES cells to selectively differentiate into primitive endoderm. Although Nanog repression occurs at the outermost layer of ES cell aggregates independent of the leukemia inhibitory factor (LIF)/STAT3 pathway, it is largely undetermined what external cues and intracellular signals cause the event. Of interest, addition of the tyrosine phosphatase inhibitor, sodium vanadate, selectively repressed Nanog transcription without any detectable changes in upstream transcriptional regulators Oct3/4 and Sox2. Furthermore, sodium vanadate induced primitive endoderm differentiation, even in the inner cells of ES cell aggregates. Expression of Gata6 and Zfp42, two putative downstream Nanog effectors, was also increased and decreased by the addition of sodium vanadate, respectively, but these changes were eliminated by exogenous Nanog expression. The effects of sodium vanadate were abrogated by Grb2 deficiency or by the addition of the Mek inhibitor, PD98059. Indeed, PD98059 prevented Nanog repression induced by ES cell aggregation as well. Furthermore, transfection of a constitutive active Mek mutant into ES cells induced Nanog repression and primitive endoderm differentiation. These data indicate that the Grb2/Mek pathway primarily mediates Nanog gene repression upon ES cell differentiation into primitive endoderm.

scholarly journals Alteration of Differentiation Potentials by Modulating GATA Transcription Factors in Murine Embryonic Stem Cells

2010 ◽  
Vol 2010 ◽  
pp. 1-15 ◽  
Author(s):  
Callinice D. Capo-chichi ◽  
Jennifer L. Smedberg ◽  
Malgorzata Rula ◽  
Emmanuelle Nicolas ◽  
Anthony T. Yeung ◽  
...  
Keyword(s):  
Es Cells ◽  
Embryonic Stem ◽  
Gene Expression Pattern ◽  
Cell Types ◽  
Primitive Endoderm ◽  
Gata Factors ◽  
Endoderm Differentiation ◽  
Collagen Iii ◽  
Hepatocyte Nuclear Factor 4

Background. Mouse embryonic stem (ES) cells can be differentiated in vitro by aggregation and/or retinoic acid (RA) treatment. The principal differentiation lineage in vitro is extraembryonic primitive endoderm. Dab2, Laminin, GATA4, GATA5, and GATA6 are expressed in embryonic primitive endoderm and play critical roles in its lineage commitment.Results. We found that in the absence of GATA4 or GATA5, RA-induced primitive endoderm differentiation of ES cells was reduced. GATA4 (−/−) ES cells express higher level of GATA5, GATA6, and hepatocyte nuclear factor 4 alpha marker of visceral endoderm lineage. GATA5 (−/−) ES cells express higher level of alpha fetoprotein marker of early liver development. GATA6 (−/−) ES cells express higher level of GATA5 as well as mesoderm and cardiomyocyte markers which are collagen III alpha-1 and tropomyosin1 alpha. Thus, deletion of GATA6 precluded endoderm differentiation but promoted mesoderm lineages.Conclusions. GATA4, GATA5, and GATA6 each convey a unique gene expression pattern and influences ES cell differentiation. We showed that ES cells can be directed to avoid differentiating into primitive endoderm and to adopt unique lineages in vitro by modulating GATA factors. The finding offers a potential approach to produce desirable cell types from ES cells, useful for regenerative cell therapy.

scholarly journals Embryonic stem cells express multiple Eph-subfamily receptor tyrosine kinases.

1996 ◽  
Vol 93 (1) ◽  
pp. 145-150 ◽  
Author(s):  
J. D. Lickliter ◽  
F. M. Smith ◽  
J. E. Olsson ◽  
K. L. Mackwell ◽  
A. W. Boyd
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Tyrosine Kinases ◽  
Receptor Tyrosine Kinases ◽  
Embryonic Stem
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