24 research outputs found
Nuclear positioning rather than contraction controls ordered rearrangements of immunoglobulin loci
Progenitor-B cells recombine their immunoglobulin (Ig) loci to create unique antigen receptors. Despite a common recombination machinery, the Ig heavy and Ig light chain loci rearrange in a stepwise manner. We studied pre-pro-B cells and Rag-/- progenitor-B cells to determine whether Ig locus contraction or nuclear positioning is decisive for stepwise rearrangements. We found that both Ig loci were contracted in pro-B and pre-B cells. Igh relocated from the nuclear lamina to central domains only at the pro-B cell stage, whereas, Igê remained sequestered at the lamina, and only at the pre-B cell stage located to central nuclear domains. Finally, in vitro induced re-positioning of Ig alleles away from the nuclear periphery increased germline transcription of Ig loci in pre-pro-B cells. Thus, Ig locus contraction juxtaposes genomically distant elements to mediate efficient recombination, however, sequential positioning of Ig loci away from the nuclear periphery determines stage-specific accessibility of Ig loci
The 3D-structure of the Immunoglobulin Heavy Chain Locus: implications for long-range genomic interactions [supplemental data]
The immunoglobulin heavy-chain (Igh) locus is organized
into distinct regions that contain multiple variable
(VH), diversity (DH), joining (JH) and constant
(CH) coding elements. How the Igh locus is structured
in 3D space is unknown. To probe the topography of
the Igh locus, spatial distance distributions were determined
between 12 genomic markers that span the
entire Igh locus. Comparison of the distance distributions
to computer simulations of alternative chromatin
arrangements predicted that the Igh locus is
Analysis of prerequisites violations financial stability
Світова економічна криза 2007–2008 років і потрясіння, що охо-
пили одночасно секторальні ринки кредитування, страхування, нерухомості та цінних паперів, продемонстрували, що системні ризики
підтримки фінансової стабільності не були належним чином оцінені
регуляторами
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified
Complementary Proteomic Tools for the Dissection of Apoptotic Proteolysis Events
Proteolysis is a key regulatory event that controls intracellular
and extracellular signaling through irreversible changes in a protein’s
structure that greatly alters its function. Here we describe a platform
for profiling caspase substrates which encompasses two highly complementary
proteomic techniquesthe first is a differential gel based
approach termed Global Analyzer of SILAC-derived Substrates of Proteolysis
(GASSP) and the second involves affinity enrichment of peptides containing
a C-terminal aspartic acid residue. In combination, these techniques
have enabled the profiling of a large cellular pool of apoptotic-mediated
proteolytic events across a wide dynamic range. By applying this integrated
proteomic work flow to analyze proteolytic events resulting from the
induction of intrinsic apoptosis in Jurkat cells via etoposide treatment,
3346 proteins were quantified, of which 360 proteins were identified
as etoposide-induced proteolytic substrates, including 160 previously
assigned caspase substrates. In addition to global profiling, a targeted
approach using BAX HCT116 isogenic cell lines was utilized to dissect
pre- and post-mitochondrial extrinsic apoptotic cleavage events. By
employing apoptotic activation with a pro-apoptotic receptor agonist
(PARA), a limited set of apoptotic substrates including known caspase
substrates such as BH3 interacting-domain death agonist (BID) and
Poly (ADP-ribose) polymerase (PARP)-1, and novel substrates such as
Basic Transcription Factor 3, TRK-fused gene protein (TFG), and p62/Sequestosome
were also identified