Transcriptome analysis reveals a ribosome constituents disorder involved in the RPL5 downregulated zebrafish model of Diamond-Blackfan anemia.

Yang Wan,Tao Cheng,Weiping Yuan,Zhaojun Zhang,Qiong Jia,Anskar Yu-Hung Leung,Xiangdong Fang,Qian Zhang,Yingchi Zhang,Haibo Jia,Xiaofan Zhu,Xiaomin Wang,Binfeng Song

doi:10.1186/s12920-016-0174-9

Abstract

BackgroundDiamond–Blackfan anemia (DBA) was the first ribosomopathy associated with mutations in ribosome protein (RP) genes. The clinical phenotypes of DBA include failure of erythropoiesis, congenital anomalies and cancer predisposition. Mutations in RPL5 are reported in approximately 9 ~ 21 % of DBA patients, which represents the most common pathological condition related to a large-subunit ribosomal protein. However, it remains unclear how RPL5 downregulation results in severe phenotypes of this disease.ResultsIn this study, we generated a zebrafish model of DBA with RPL5 morphants and implemented high-throughput RNA-seq and ncRNA-seq to identify key genes, lncRNAs, and miRNAs during zebrafish development and hematopoiesis. We demonstrated that RPL5 is required for both primitive and definitive hematopoiesis processes. By comparing with other DBA zebrafish models and processing functional coupling network, we identified some common regulated genes, lncRNAs and miRNAs, that might play important roles in development and hematopoiesis.ConclusionsRibosome biogenesis and translation process were affected more in RPL5 MO than in other RP MOs. Both P53 dependent (for example, cell cycle pathway) and independent pathways (such as Aminoacyl-tRNA biosynthesis pathway) play important roles in DBA pathology. Our results therefore provide a comprehensive basis for the study of molecular pathogenesis of RPL5-mediated DBA and other ribosomopathies.Electronic supplementary materialThe online version of this article (doi:10.1186/s12920-016-0174-9) contains supplementary material, which is available to authorized users.

Highlights

Diamond–Blackfan anemia (DBA) was the first ribosomopathy associated with mutations in ribosome protein (RP) genes
Hematopoiesis and morphological abnormalities in Ribosomal protein L5 (RPL5) downregulation zebrafish There are two isoforms of RPL5 (RPL5a and RPL5b), and the ATG MO that we designed could inhibit both isoforms in vivo
The effectiveness of the translation inhibition by RPL5 morpholino (RPL5 MO) was confirmed by examining green fluorescent fusion protein expression by fluorescence microscopy (Fig. 1a and b)

Summary

Introduction

Diamond–Blackfan anemia (DBA) was the first ribosomopathy associated with mutations in ribosome protein (RP) genes. Mutations in RPL5 are reported in approximately 9 ~ 21 % of DBA patients, which represents the most common pathological condition related to a large-subunit ribosomal protein. It remains unclear how RPL5 downregulation results in severe phenotypes of this disease. Diamond-Blackfan anemia (DBA) (MIM 105650) was the first ribosomopathy to be associated with genetic mutations in ribosomal proteins when a mutation in ribosomal protein S19 (RPS19) was first reported in 1999 [2] It is known as a rare inherited bone marrow failure syndromes (IBMFS) characterized by the failure of erythropoiesis with normal platelet and myeloid lineages. Since the initial identification of RPS19, approximately 50– 60 % of DBA patients have been found to have mutations or deletions in genes encoding ribosomal protein (RP) of both the small and large subunits, including RPS24, RPS17, RPL35A, RPL5, RPL11, RPS7, RPS10, RPS26, RPL26 and RPS29 [6, 7]

Methods

Results

Conclusion