Characterization of Histone Genes from the Bivalve Lucina Pectinata.

Ingrid Montes-Rodríguez,Carmen Cadilla,Alexander Ropelewski,Juan Lopez-Garriga,Yesenia Rodríguez-Pou,Ricardo González-Méndez

doi:10.3390/ijerph15102170

Abstract

Lucina pectinata is a clam that lives in sulfide-rich environments and houses intracellular sulfide-oxidizing endosymbionts. To identify new Lucina pectinata proteins, we produced libraries for genome and transcriptome sequencing and assembled them de novo. We searched for histone-like sequences using the Lucina pectinata histone H3 partial nucleotide sequence against our previously described genome assembly to obtain the complete coding region and identify H3 coding sequences from mollusk sequences in Genbank. Solen marginatus histone nucleotide sequences were used as query sequences using the genome and transcriptome assemblies to identify the Lucina pectinata H1, H2A, H2B and H4 genes and mRNAs and obtained the complete coding regions of the five histone genes by RT-PCR combined with automated Sanger DNA sequencing. The amino acid sequence conservation between the Lucina pectinata and Solen marginatus histones was: 77%, 93%, 83%, 96% and 97% for H1, H2A, H2B, H3 and H4, respectively. As expected, the H3 and H4 proteins were the most conserved and the H1 proteins were most similar to H1′s from aquatic organisms like Crassostrea gigas, Aplysia californica, Mytilus trossulus and Biomphalaria glabrata. The Lucina pectinata draft genome and transcriptome assemblies, obtained by semiconductor sequencing, were adequate for identification of conserved proteins as evidenced by our results for the histone genes.

Highlights

In eukaryotes and some archaebacteria, DNA is found as a nucleoprotein complex called chromatin, associated with histones that allows for the high levels of compaction of genomicDNA within the limited space of the cell nucleus [1]
Our strategy to search for histone genes relied on the fact that histones have been highly conserved in evolution
histone 4 (H4) and histone 3 (H3) are highly conserved while histone 1 (H1), histone 2A (H2A) and histone 2B (H2B) are less conserved [21]

Summary

Introduction

In eukaryotes and some archaebacteria, DNA is found as a nucleoprotein complex called chromatin, associated with histones that allows for the high levels of compaction of genomic. DNA within the limited space of the cell nucleus [1]. Four of them (H3, H4, H2A and H2B) form the histone octamer that DNA winds around to form the nucleosome, the fundamental structural unit of chromatin [2]. A fifth lysine-rich histone (H1 and related proteins) binds the nucleosome at the entry and exit sites of the DNA, locking the DNA into place and allowing the development of higher order structures. Histones play a major role in modulating changes in chromatin structure, impacting gene regulation, mostly through their post-translational modifications [3]. Public Health 2018, 15, 2170; doi:10.3390/ijerph15102170 www.mdpi.com/journal/ijerph

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