The exosome complex establishes a barricade to erythroid maturation

Slides:



Advertisements
Similar presentations
GCN5 Regulates FGF Signaling and Activates Selective MYC Target Genes during Early Embryoid Body Differentiation  Li Wang, Evangelia Koutelou, Calley.
Advertisements

STAT3 mediates oncogenic addiction to TEL-AML1 in t(12;21) acute lymphoblastic leukemia by Maurizio Mangolini, Jasper de Boer, Vanessa Walf-Vorderwülbecke,
by Elena A. Federzoni, Peter J. M. Valk, Bruce E
Quantitative analysis of murine terminal erythroid differentiation in vivo: novel method to study normal and disordered erythropoiesis by Jing Liu, Jianhua.
Identification of key regulatory pathways of myeloid differentiation using an mESC-based karyotypically normal cell model by Dong Li, Hong Yang, Hong Nan,
HOXA9 promotes hematopoietic commitment of human embryonic stem cells
by Leah J. Anderson, and Richard Longnecker
The PP2A inhibitor SET regulates granzyme B expression in human natural killer cells by Rossana Trotta, David Ciarlariello, Jessica Dal Col, Hsiaoyin Mao,
by Sang-Hyun Song, AeRi Kim, Tobias Ragoczy, M. A
by Susan E. Shetzline, Ravikumar Rallapalli, Kelley J
by Shawn W. Cochrane, Ying Zhao, Robert S. Welner, and Xiao-Hong Sun
Transcription factor GATA-1 potently represses the expression of the HIV-1 coreceptor CCR5 in human T cells and dendritic cells by Mark S. Sundrud, Scott.
The C/EBPδ tumor suppressor is silenced by hypermethylation in acute myeloid leukemia by Shuchi Agrawal, Wolf-Karsten Hofmann, Nicola Tidow, Mathias Ehrich,
by Andrew G. Muntean, Liyan Pang, Mortimer Poncz, Steven F
EHMT1 and EHMT2 inhibition induces fetal hemoglobin expression
FOG-1 represses GATA-1-dependent FcϵRI β-chain transcription: transcriptional mechanism of mast-cell-specific gene expression in mice by Keiko Maeda, Chiharu.
by Kevin Oakley, Yufen Han, Bandana A
by Silke Huber, Reinhard Hoffmann, Femke Muskens, and David Voehringer
Activation of ATF4 mediates unwanted Mcl-1 accumulation by proteasome inhibition by Jinsong Hu, Nana Dang, Eline Menu, Elke De Bryune, Dehui Xu, Ben Van.
by Jessica M. Salmon, Nicholas J. Slater, Mark A. Hall, Matthew P
Activation of the vitamin D receptor transcription factor stimulates the growth of definitive erythroid progenitors by Jeffrey Barminko, Brad M. Reinholt,
Bone morphogenetic protein 4 induces efficient hematopoietic differentiation of rhesus monkey embryonic stem cells in vitro by Fei Li, Shijiang Lu, Loyda.
Volume 19, Issue 12, Pages (June 2017)
ICSBP/IRF-8 inhibits mitogenic activity of p210 Bcr/Abl in differentiating myeloid progenitor cells by Tomohiko Tamura, Hee Jeong Kong, Chainarong Tunyaplin,
Suppression of Fas-FasL coexpression by erythropoietin mediates erythroblast expansion during the erythropoietic stress response in vivo by Ying Liu, Ramona.
Developmental- and differentiation-specific patterns of human γ- and β-globin promoter DNA methylation by Rodwell Mabaera, Christine A. Richardson, Kristin.
Cooperative signaling between cytokine receptors and the glucocorticoid receptor in the expansion of erythroid progenitors: molecular analysis by expression.
by Anupama Narla, Shilpee Dutt, J
The transcriptional program of terminal granulocytic differentiation
Therapeutic levels of fetal hemoglobin in erythroid progeny of β-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer by Andrew Wilber,
Elucidation of the EP defect in Diamond-Blackfan anemia by characterization and prospective isolation of human EPs by Deena Iskander, Bethan Psaila, Gareth.
by Guang Yang, Shu-Ching Huang, Jane Y. Wu, and Edward J. Benz
BET inhibition and depletion repress the expression of BRCA1 and RAD51
LEF-1 is a prosurvival factor in chronic lymphocytic leukemia and is expressed in the preleukemic state of monoclonal B-cell lymphocytosis by Albert Gutierrez,
Identification and characterization of 2 types of erythroid progenitors that express GATA-1 at distinct levels by Norio Suzuki, Naruyoshi Suwabe, Osamu.
Volume 6, Issue 3, Pages (March 2016)
Volume 16, Issue 9, Pages (August 2016)
by Gregory H. Underhill, David George, Eric G. Bremer, and Geoffrey S
Lentiviral-mediated RNAi inhibition of Sbds in murine hematopoietic progenitors impairs their hematopoietic potential by Amy S. Rawls, Alyssa D. Gregory,
Soluble PD-1 ligands regulate T-cell function in Waldenstrom macroglobulinemia by Shahrzad Jalali, Tammy Price-Troska, Jonas Paludo, Jose Villasboas, Hyo-Jin.
Bmi-1 Regulates Extensive Erythroid Self-Renewal
by Kentson Lam, Alexander Muselman, Randal Du, Yuka Harada, Amanda G
Volume 31, Issue 4, Pages (October 2009)
MiR-34a contributes to megakaryocytic differentiation of K562 cells independently of p53 by Francisco Navarro, David Gutman, Eti Meire, Mario Cáceres,
Dynamic change of transcription pausing through modulating NELF protein stability regulates granulocytic differentiation by Xiuli Liu, Aishwarya A. Gogate,
Wenqian Hu, Bingbing Yuan, Harvey F. Lodish  Developmental Cell 
Induced pluripotent stem cell–based mapping of β-globin expression throughout human erythropoietic development by Kim Vanuytsel, Taylor Matte, Amy Leung,
Volume 9, Issue 3, Pages (September 2017)
Volume 19, Issue 12, Pages (June 2017)
Sphingosine-1-phosphate signaling modulates terminal erythroid differentiation through the regulation of mitophagy  Chong Yang, Michihiro Hashimoto, Quy.
Volume 10, Issue 1, Pages (January 2018)
Figure 1. BRCA1-associated R-Loop accumulation at a non-coding region upstream of ESR1 locus. (A) Alignment of DRIP-seq ... Figure 1. BRCA1-associated.
Volume 20, Issue 12, Pages (September 2017)
Myeloma cell–derived Runx2 promotes myeloma progression in bone
Volume 158, Issue 4, Pages (August 2014)
A Genetic Screen Identifies TCF3/E2A and TRIAP1 as Pathway-Specific Regulators of the Cellular Response to p53 Activation  Zdenek Andrysik, Jihye Kim,
by Vijay G. Sankaran, Tobias F. Menne, Jian Xu, Thomas E
Twist1 regulates embryonic hematopoietic differentiation through binding to Myb and Gata2 promoter regions by Kasem Kulkeaw, Tomoko Inoue, Tadafumi Iino,
Volume 3, Issue 4, Pages (October 2008)
AZA treatment induces a distinct gene-expression pattern in stromal cells. AZA treatment induces a distinct gene-expression pattern in stromal cells. (A-C)
P300 depletion is lethal in cancer cells harboring loss-of-function mutations in CBP. A, synthetic-lethal effects assessed by colony formation assay. p300.
Volume 23, Issue 8, Pages (May 2018)
Volume 17, Issue 4, Pages (October 2015)
Fig. 7. SCF and IGF-1 accelerate fetal spleen erythropoiesis in vivo.
Volume 16, Issue 2, Pages (February 2002)
Silencing is not established at a hybrid HML-Pa locus without passage through S phase. Silencing is not established at a hybrid HML-Pa locus without passage.
Volume 11, Issue 21, Pages (October 2001)
Volume 21, Issue 9, Pages (November 2017)
Volume 33, Issue 1, Pages (July 2010)
Volume 12, Issue 2, Pages (February 2013)
Presentation transcript:

The exosome complex establishes a barricade to erythroid maturation by Skye C. McIver, Yoon-A Kang, Andrew W. DeVilbiss, Chelsea A. O’Driscoll, Jonathan N. Ouellette, Nathaniel J. Pope, Genis Camprecios, Chan-Jung Chang, David Yang, Eric E. Bouhassira, Saghi Ghaffari, and Emery H. Bresnick Blood Volume 124(14):2285-2297 October 2, 2014 ©2014 by American Society of Hematology

GATA-1/Foxo3-dependent genetic network. GATA-1/Foxo3-dependent genetic network. (A) Venn diagrams depicting genes regulated uniquely or coregulated by GATA-1 and Foxo3. (B) Venn diagrams demonstrating relationships between GATA-1– and Foxo3-activated and -repressed genes. (C) Gene ontology analysis of GATA-1/Foxo3-coactivated genes. (D) Gene ontology analysis of GATA-1– and Foxo3-corepressed genes. The top 10 gene ontology categories are displayed and ordered by P value. Redundant gene ontology categories were curated and removed. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

GATA-1/Foxo3-mediated repression of Exosc8 expression. GATA-1/Foxo3-mediated repression of Exosc8 expression. (A) Real-time RT-PCR analysis of Foxo3, Exosc8 mRNA and primary transcripts upon GATA-1 activation in G1E-ER-GATA-1 cells (mean ± SE; 3 independent experiments). Values were normalized to 18S rRNA expression. (B) Semiquantitative western blot analysis of Foxo3 in G1E-ER-GATA-1 cells after 0, 24, or 48 hours estradiol treatment (left). The representative image is shown from 3 independent experiments. Quantitation of band intensity (mean ± SE; 3 independent experiments) (right). (C) Real-time RT-PCR analysis of Foxo3 and Exosc8 mRNA upon Foxo3 knockdown in G1E-ER-GATA-1 cells after 42 hours activation of GATA-1 (mean ± standard error [SE]; 4 independent experiments). Values were normalized to 18S rRNA expression and expression is shown relative to the control. (D) ChIP-seq profile of GATA-1 occupancy at EXOSC8 in primary human erythroblasts. (E) ChIP analysis of Foxo3 occupancy at the Exosc8 promoter in untreated and estradiol-treated (24 hours) G1E-ER-GATA-1 cells (mean ± SE; 4 independent experiments). (F) Real-time RT-PCR analysis of mRNA levels in control vs Exosc8 knockdown G1E-ER-GATA-1 cells (mean ± SE; 3 independent experiments). Values were normalized to 18S rRNA expression. mRNA levels are shown relative to the control siRNA with estradiol treatment of activated genes (left) and without estradiol treatment of repressed genes (middle). Fold changes upon Exosc8 knockdown relative to control (right). (G) Real-time RT-PCR analysis of primary transcripts in control vs Exosc8 knockdown G1E-ER-GATA-1 cells (mean ± SE; 3 independent experiments). Values were normalized to 18S rRNA expression, and the expression is shown relative to estradiol-treated control siRNA. Fold changes are also depicted upon Exosc8 knockdown relative to control (right). *P < .05, **P < .01, ***P < .001. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

Repression of exosome-complex component expression during erythroid maturation. Repression of exosome-complex component expression during erythroid maturation. (A) Schematic diagram and crystal structure of the human exosome complex.26 Solid line, direct interactions; dashed line, indirect interactions; red, associated components with catalytic activity. (B) Expression profile of mRNA levels for exosome complex components in primary mouse bone marrow erythroblasts during distinct maturation stages mined from the Erythron DB.29 P, proerythroblast; B, basophilic erythroblast; O, orthochromatic erythroblast; R, reticulocyte. (C) Expression profile of mRNAs encoding exosome-complex components during primary human erythroid differentiation mined from the Human Erythroblast Maturation (HEM) Database.30 C, colony-forming unit-erythroid (CFU-E); P, proerythroblast; I, intermediate-stage erythroblast; L, late-stage erythroblast. (D) Expression profile of mRNAs encoding exosome-complex components during ex vivo differentiation of primary mouse fetal liver–derived erythroid precursor cells. 1, R1 population: progenitor cells; 2, R2 population: proerythroblasts and early basophilic erythroblasts; 3, R3 population: early and late basophilic erythroblasts. 4, R4/5 population: polychromatophilic, orthochromatic erythroblasts, and reticulocytes. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

Exosc8-dependent barrier to erythroid maturation. Exosc8-dependent barrier to erythroid maturation. (A) Real-time RT-PCR analysis of Exosc8 and selected GATA-1 target gene mRNA levels in control vs Exosc8 knockdown primary murine erythroid precursor cells cultured under expansion (–) or differentiation conditions (+) (mean ± SE; 3 independent experiments). Values were normalized to 18S rRNA expression and the expression is shown relative to control shRNA under expansion conditions. (B) Flow cytometric quantitation of erythroid developmental stage by CD71 and Ter119 staining upon Exosc8 knockdown in primary erythroid precursor cells. Representative flow cytometry data, with the R1-R5 gates denoted from 3 independent experiments. The percentage of live cells from each condition and the cell populations in R1-R5 stages (mean ± SE; 3 independent experiments). E, expansion; D, differentiation. (C) Representative images of Wright-Giemsa staining in control vs Exosc8 shRNA–infected primary erythroid precursor cells cultured in expansion or differentiation media (scale bar = 10 µm) and quantitation of cell size by measuring forward scatter using flow cytometry. (D) Flow cytometric cell-cycle analysis of primary erythroid precursor cells infected with retrovirus expressing control or Exosc8 2 shRNA. Representative cell-cycle profile is shown from 3 independent experiments. The percentage of the cell population in each cell-cycle stage is from 3 independent experiments (mean ± SE). Blue, S phase; red, G0/G1 or G2/M phase. (E) Quantitative ChIP analysis of serine 5-phosphorylated RNA Polymerase II occupancy at Exosc8-regulated GATA-1 target and control genes in control and Exosc8-knockdown primary murine erythroid precursor cells (mean ± SE; 3 independent experiments). *P < .05, **P < .01, ***P < .001. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

Evidence that the exosome complex creates a barrier to erythroid maturation. Evidence that the exosome complex creates a barrier to erythroid maturation. (A) Crystal structure of the human exosome complex26 demonstrating the interaction between Exosc8 and Exosc9. (B) Real-time RT-PCR analysis of Exosc8 and Exosc9 mRNA levels in control vs Exosc8 or Exosc9 knockdown in expanding primary murine erythroid precursor cells sorted into distinct, R1, R2, R3, and R4/5 cell populations (mean ± SE; 5 independent experiments). Values were normalized to 18S rRNA level and the expression relative to the control R1 shRNA condition. (C) Semiquantitative western blot analysis of Exosc9 protein in expanding primary murine erythroid precursor cells 24 hours postinfection with Exosc8- or Exosc9-specific shRNA retroviruses. Representative image from 3 independent experiments. (D) Real-time RT-PCR analysis of selected GATA-1 target genes and erythroid transcription factors mRNA levels in control vs Exosc8 or Exosc9 knockdown in expanding primary murine erythroid precursor cells sorted into distinct, R1, R2, R3, and R4/5 cell populations (mean ± SE; 5 independent experiments). Values were normalized to 18S rRNA level and the expression relative to the control R1 shRNA condition. (E) Flow cytometric quantitation of erythroid maturation stage by CD71 and Ter119 staining upon Exosc8 or Exosc9 knockdown in primary erythroid precursor cells. Representative flow cytometry data, with the R1-R5 gates denoted from 4 independent experiments. The percentage of the cell populations in R1-R5 stages (mean ± SE; 4 independent experiments). (F) Representative images of Wright-Giemsa–stained cells infected with control vs Exosc8 or Exosc9 shRNA-expressing virus. Cells were cultured under expansion conditions (scale bar = 10 µm). (G) Cell number fold change during the 72-hour expansion of primary murine erythroid precursor cells postinfection with control, Exosc8, or Exosc9 shRNA-expressing retroviruses (mean ± SE; 8 independent experiments). (H) Flow cytometric quantitation of erythroid maturation stage by CD71 and Ter119 staining upon Exosc8 or Dis3 knockdown in primary erythroid precursor cells. Representative flow cytometry data, with the R1-R5 gates denoted. The percentage of the cell populations in R1-R5 stages (mean ± SE; 6 independent experiments. *P < .05, **P < .01, ***P < .001. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

GATA-1/Foxo3/Exosc8 coregulate genes important for erythroid maturation. GATA-1/Foxo3/Exosc8 coregulate genes important for erythroid maturation. (A) Classification of GATA-1– and Exosc8-regulated genes based on activation or repression. (B) Gene ontology analysis of GATA-1–activated, Exosc8-repressed genes. Redundant Gene Ontology categories were curated and removed. (C) Classification of Foxo3- and Exosc8-regulated genes based on activation or repression. (D) Gene ontology analysis of Foxo3-activated, Exosc8-repressed genes. Redundant Gene Ontology categories were curated and removed. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology

The exosome complex suppresses cell-cycle arrest genes during erythroid maturation. The exosome complex suppresses cell-cycle arrest genes during erythroid maturation. (A) Name and function41-43,45,47,52,61 of genes in “cell cycle arrest” category derived from GO term analysis from GATA-1–activated and Exosc8-repressed genes. (B) ChIP-seq profiles of GATA-1 occupancy at cell-cycle regulatory genes in primary human erythroblasts. All genes were orientated left to right. (C) Real-time RT-PCR analysis of genes in “cell cycle arrest” category upon Exosc8 or Exosc9 knockdown in primary murine erythroid precursor cells under expansion conditions, sorted into distinct, R1, R2, R3, and R4/5 cell populations (mean ± SE; 5 independent experiments). Values were normalized to 18S rRNA and the expression relative to the control R1 population. (D) Flow cytometric cell cycle analysis of primary erythroid precursor cells, within the R3 population, infected with retrovirus expressing control, Exosc8, or Exosc9 shRNA. Representative cell-cycle profile from 2 independent experiments. The percentage of the cell population in each cell cycle stage (mean ± SE; 2 independent experiments). Shaded, S phase; red, G0/G1 or G2/M phase. *P < .05, **P < .01, ***P < .001. (E) Flow cytometric cell cycle analysis of primary erythroid precursor cells treated with 25 μM HU for either 24 or 48 hours. Representative cell-cycle profile. The percentage of the cell population in each cell cycle stage (mean ± SE; 3 independent experiments). Blue, S phase; red, G0/G1 or G2/M phase. (F) Flow cytometric quantitation of erythroid maturation stage by CD71 and Ter119 staining upon 25 μM HU treatment of 24 or 48 hours in primary erythroid precursor cells. Representative flow cytometry data, with the R1-R5 gates denoted (3 independent experiments). (G) Model of GATA-1/Foxo3 function to overcome the exosome complex–dependent erythroid maturation barricade, which involves multiple alterations in erythroid cell function. Skye C. McIver et al. Blood 2014;124:2285-2297 ©2014 by American Society of Hematology