Characterisation of functional domains in fission yeast Ams2 that are required for core histone gene transcription.

Yuko Takayama,Masaki Shirai,Fumie Masuda

doi:10.1038/srep38111

Yuko Takayama, Masaki Shirai + Show 1 more

Open Access

https://doi.org/10.1038/srep38111

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Journal: Scientific reports	Publication Date: Nov 30, 2016
Citations: 5	License type: CC BY 4.0

Affiliation: Teikyo University, Kurume University

Abstract

Histone gene expression is regulated in a cell cycle-dependent manner, with a peak at S phase, which is crucial for cell division and genome integrity. However, the detailed mechanisms by which expression of histone genes are tightly regulated remain largely unknown. Fission yeast Ams2, a GATA-type zinc finger motif-containing factor, is required for activation of S phase-specific core histone gene transcription. Here we report the molecular characterisation of Ams2. We show that the zinc finger motif in Ams2 is necessary to bind the histone gene promoter region and to activate histone gene transcription. An N-terminal region of Ams2 acts as a self-interaction domain. Intriguingly, N-terminally truncated Ams2 binds to the histone gene promoters, but does not fully activate histone gene transcription. These observations imply that Ams2 self-interactions are required for efficient core histone gene transcription. Moreover, we show that Ams2 interacts with Teb1, which itself binds to the core histone gene promoters. We discuss the relationships between Ams2 domains and efficient transcription of the core histone genes in fission yeast.

Highlights

Observations, it has been proposed that Spt[10] proteins containing NTD and DBD form dimers via an N-terminal domain that stably binds to paired UAS elements[7,16]
We constructed strains in which genes encoding HA-tagged versions of Ams[2] with or without the mutated zinc finger (MZF) were separately integrated at the chromosomal ams2+ locus, permitting expression of the tagged proteins from the native promoter. These cells were synchronised in S phase by HU treatment, and the cell lysates were immunoprecipitated with anti-HA antibody
We did not detect association of the zinc finger-mutated Ams[2] with the SPAC631.02+ gene promoter (Fig. 1b, 631.02+). These results clearly showed that the zinc finger motif of Ams[2] is required for binding to the AACCT-box, which is contained in all core histone gene promoters

Summary

Introduction

Observations, it has been proposed that Spt[10] proteins containing NTD and DBD form dimers via an N-terminal domain that stably binds to paired UAS elements[7,16]. Ams[2], a GATA-type zinc finger protein, promotes the centromere localisation of the centromere-specific histone H3 variant CENP-A17. Ams[2] is required for transcriptional activation of the core histone genes at S phase, and binds to the histone gene promoter regions in vivo in an AACCCT-box dependent manner[12]. Ams[2] protein levels, which are controlled by the ubiquitin proteasome pathways[18,19], increase during the cell cycle to a peak in the G1/S phase[12,17] that ensures S phase-specific transcriptional activation of core histone genes. We examined the roles of the zinc finger motif and N-terminal domains of Ams[2] in core histone gene transcription

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Conclusion