Abstract

Drosophila underreplicate the DNA of thoracic nuclei, stalling during S phase at a point that is proportional to the total genome size in each species. In polytene tissues, such as the Drosophila salivary glands, all of the nuclei initiate multiple rounds of DNA synthesis and underreplicate. Yet, only half of the nuclei isolated from the thorax stall; the other half do not initiate S phase. Our question was, why half? To address this question, we use flow cytometry to compare underreplication phenotypes between thoracic tissues. When individual thoracic tissues are dissected and the proportion of stalled DNA synthesis is scored in each tissue type, we find that underreplication occurs in the indirect flight muscle, with the majority of underreplicated nuclei in the dorsal longitudinal muscles (DLM). Half of the DNA in the DLM nuclei stall at S phase between the unreplicated G0 and fully replicated G1. The dorsal ventral flight muscle provides the other source of underreplication, and yet, there, the replication stall point is earlier (less DNA replicated), and the endocycle is initiated. The differences in underreplication and ploidy in the indirect flight muscles provide a new tool to study heterochromatin, underreplication and endocycle control.

Highlights

  • While a species is known to have a basal ploidy level, with only the chromosome number from parents, individual tissue types and cells may differ in their copies of the genome or portions of the genome

  • We find that UR in the thorax occurs primarily, if not exclusively, in the indirect flight muscles of both species, with significant differences between the dorsal longitudinal muscle (DLM) and dorsal ventral muscle (DVM)

  • UR in the DLM

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Summary

Introduction

While a species is known to have a basal ploidy level, with only the chromosome number from parents, individual tissue types and cells may differ in their copies of the genome or portions of the genome. Found in specialty tissues of Diptera, such as the well-studied polyploid salivary gland and ovarian follicle cells, UR results from a coupling of protein products from Rif and SuUR hindering progression of the DNA replication fork [3,4]. This stall usually occurs upon encountering heterochromatic DNA, which is tightly wound into nucleosomes, preventing this heterochromatic DNA from fully replicating [5,6]. UR does not alter origin firing, or the initiation of replication [9], Genes 2020, 11, 246; doi:10.3390/genes11030246 www.mdpi.com/journal/genes

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