Abstract

Localized maternal mRNAs play important roles in embryogenesis, e.g. the establishment of embryonic axes and the developmental cell fate specification, in various animal species. In ascidians, a group of maternal mRNAs, called postplasmic/PEM RNAs, is localized to a subcellular structure, called the Centrosome-Attracting Body (CAB), which contains the ascidian germ plasm, and is inherited by the germline cells during embryogenesis. Posterior end mark (Pem), a postplasmic/PEM RNAs member, represses somatic gene expression in the germline during cleavage stages by inhibition of RNA polymerase II activity. However, the functions of other postplasmic/ PEM RNAs members in germline formation are largely unknown. In this study, we analyzed the functions of two postplasmic/PEM RNAs, Popk-1 and Zf-1, in transcriptional regulation in the germline cells. We show that Popk-1 contributes to transcriptional quiescence by controlling the size of the CAB and amount of Pem protein translated at the CAB. Our studies also indicated that zygotic expression of a germline gene starts around the onset of gastrulation and that the decrease of Pem protein is necessary and sufficient for the zygotic germline gene expression. Finally, further studies showed that the decrease of the Pem protein level is facilitated by Zf-1. Taken together, we propose that postplasmic/PEM RNAs such as Popk-1 and Zf-1 control the protein level of the transcriptional repressor Pem and regulate its transcriptional state in the ascidian germline.

Highlights

  • Germline is a specialized cellular lineage that transmits genetic information to the generation

  • In the process of searching for other factors in addition to Posterior end mark (Pem) that regulate transcriptional repression in the embryonic germline, we found that Popk-1 knockdown by morpholino antisense oligo nucleotide (MO) injection resulted in ectopic FoxD.a expression in B5.2 blastomeres, the germline cells of the 16-cell stage embryos (Fig 2A, 2B, arrows, 2E)

  • Popk-1 knockdown resulted in decrease of the Centrosome-Attracting Body (CAB) size as well as the amount of postplasmic/PEM RNAs localized to CAB [37], where translation of the localized mRNAs takes place [49]

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Summary

Introduction

Germline is a specialized cellular lineage that transmits genetic information to the generation. The germline is set aside from the somatic linage throughout their life cycles. This separation plays a pivotal role in the retention of the unique characteristics of germ cells such as their totipotency and immortality and in protection from being compromised by somatic programs. One strategy for this germline segregation is known as transcriptional repression for somatic genes in the lineage [1,2,3,4].

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