Proposed research:
Aims and Objectives:
Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development
Aim2: To characterise rDNA chromatin organisation in the oocyte
Aim3: To elucidate the transcriptional machinery of RNA Polymerase I in the oocyte
Experimental Approach:
Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development
The Hira histone chaperone complex is an evolutionarily conserved complex, which is composed of Hira, Ubn1 and Cabin1, and cooperates with the histone chaperone Asf1a to mediate H3.3-specific binding and chromatin deposition12,13. I previously demonstrated the critical roles of maternal Hira /H3.3 on paternal pronucleus formation and developmental competence using a genetically engineered mouse model. However, the maternal functions of Cabin1 and Ubn1 remain unknown. I therefore propose to characterise the functional roles of Cabin1 and Ubn1 …show more content…
RRNA is the key component of ribosomes, which are account for translation. The role of Hira and H3.3 on rRNA transcription prompts me to investigate whether global translational process is compromised. The defects on handful ribosomal protein (Rpl9 and Rpl10A) indirectly support this idea. However, only a detailed measurement of translation genome-widely will provide and reveal what are the key players that are required for developmental competence and which of these are shortage in Hira mutants. I will take risk and develop/explore the feasibility on applying Ribosome profiling technology (using ARTseq) on mouse oocytes, which have not been conducted yet. The data will reveal the profile of active translation and further integrate with total RNA-seq data will uncover the translation levels. By comparing wild type versus Hira (Cabin/Ubn1 if needed) will reveal the differences of translatome, which will underline the candidates for oocyte
Aims and Objectives:
Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development
Aim2: To characterise rDNA chromatin organisation in the oocyte
Aim3: To elucidate the transcriptional machinery of RNA Polymerase I in the oocyte
Experimental Approach:
Aim1: To address the roles of the Hira complex in rRNA transcription and embryonic development
The Hira histone chaperone complex is an evolutionarily conserved complex, which is composed of Hira, Ubn1 and Cabin1, and cooperates with the histone chaperone Asf1a to mediate H3.3-specific binding and chromatin deposition12,13. I previously demonstrated the critical roles of maternal Hira /H3.3 on paternal pronucleus formation and developmental competence using a genetically engineered mouse model. However, the maternal functions of Cabin1 and Ubn1 remain unknown. I therefore propose to characterise the functional roles of Cabin1 and Ubn1 …show more content…
RRNA is the key component of ribosomes, which are account for translation. The role of Hira and H3.3 on rRNA transcription prompts me to investigate whether global translational process is compromised. The defects on handful ribosomal protein (Rpl9 and Rpl10A) indirectly support this idea. However, only a detailed measurement of translation genome-widely will provide and reveal what are the key players that are required for developmental competence and which of these are shortage in Hira mutants. I will take risk and develop/explore the feasibility on applying Ribosome profiling technology (using ARTseq) on mouse oocytes, which have not been conducted yet. The data will reveal the profile of active translation and further integrate with total RNA-seq data will uncover the translation levels. By comparing wild type versus Hira (Cabin/Ubn1 if needed) will reveal the differences of translatome, which will underline the candidates for oocyte