Poly(C)-Binding Protein Pcbp2 Enables Differentiation of Definitive Erythropoiesis by Directing Functional Splicing of the Runx1 Transcript

Louis R Ghanem; Andrew Kromer; Ian M Silverman; Xinjun Ji; Matthew Gazzara; Nhu Nguyen; Gabrielle Aguilar; Massimo Martinelli; Yoseph Barash; Stephen A Liebhaber

doi:10.1128/MCB.00175-18

Poly(C)-Binding Protein Pcbp2 Enables Differentiation of Definitive Erythropoiesis by Directing Functional Splicing of the Runx1 Transcript

Mol Cell Biol. 2018 Jul 30;38(16):e00175-18. doi: 10.1128/MCB.00175-18. Print 2018 Aug 15.

Authors

Louis R Ghanem^{1

2

3}, Andrew Kromer³, Ian M Silverman³, Xinjun Ji³, Matthew Gazzara³, Nhu Nguyen³, Gabrielle Aguilar⁴, Massimo Martinelli^{4

5}, Yoseph Barash³, Stephen A Liebhaber^{6

7}

Affiliations

¹ Gastroenterology, Hepatology and Nutrition Division, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA ghaneml@email.chop.edu liebhabe@mail.med.upenn.edu.
² Department of Pediatrics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA.
³ Department of Genetics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA.
⁴ Gastroenterology, Hepatology and Nutrition Division, The Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA.
⁵ Department of Translational Medical Science, Section of Pediatrics, University of Naples Federico II, Naples, Italy.
⁶ Department of Genetics, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA ghaneml@email.chop.edu liebhabe@mail.med.upenn.edu.
⁷ Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Abstract

Formation of the mammalian hematopoietic system is under a complex set of developmental controls. Here, we report that mouse embryos lacking the KH domain poly(C) binding protein, Pcbp2, are selectively deficient in the definitive erythroid lineage. Compared to wild-type controls, transcript splicing analysis of the Pcbp2^-/- embryonic liver reveals accentuated exclusion of an exon (exon 6) that encodes a highly conserved transcriptional control segment of the hematopoietic master regulator, Runx1. Embryos rendered homozygous for a Runx1 locus lacking this cassette exon (Runx1ΔE6) effectively phenocopy the loss of the definitive erythroid lineage in Pcbp2^-/- embryos. These data support a model in which enhancement of Runx1 cassette exon 6 inclusion by Pcbp2 serves a critical role in development of hematopoietic progenitors and constitutes a critical step in the developmental pathway of the definitive erythropoietic lineage.

Keywords: Pcbp2; RNA processing; Runx1; alternative splicing; definitive erythropoiesis; erythropoiesis; hematopoiesis; myeloid stem cells; posttranscriptional control mechanisms; primitive erythropoiesis.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Sequence
Animals
Base Sequence
Cell Differentiation / genetics
Cell Differentiation / physiology
Core Binding Factor Alpha 2 Subunit / deficiency
Core Binding Factor Alpha 2 Subunit / genetics*
Core Binding Factor Alpha 2 Subunit / metabolism
Erythropoiesis / genetics*
Erythropoiesis / physiology*
Exons
Gene Expression Regulation, Developmental
Globins / genetics
Hematopoiesis / genetics
Hematopoiesis / physiology
Humans
K562 Cells
Liver / embryology
Liver / metabolism
Mice
Mice, Knockout
RNA Splicing
RNA-Binding Proteins / genetics*
RNA-Binding Proteins / metabolism*
Sequence Deletion

Substances

Core Binding Factor Alpha 2 Subunit
Pcbp2 protein, mouse
RNA-Binding Proteins
Runx1 protein, mouse
Globins

Abstract

Publication types

MeSH terms

Substances

Grants and funding