High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Spermatogenesis.

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RESEARCH PRODUCT

High Throughput Sequencing Identifies Misregulated Genes in the Drosophila Polypyrimidine Tract-Binding Protein (hephaestus) Mutant Defective in Spermatogenesis.

Joseph Heimiller Vinod Sridharan Mark D. Robida Ravinder Singh

subject

0301 basic medicine Male Physiology Mutant Gene Expression lcsh:Medicine Artificial Gene Amplification and Extension Polymerase Chain Reaction Biochemistry Conserved sequence 0302 clinical medicine Sequencing techniques Reproductive Physiology Animal Cells Invertebrate Genomics Medicine and Health Sciences Drosophila Proteins Protein Isoforms Cell Cycle and Cell Division lcsh:Science Conserved Sequence Phylogeny Genetics Regulation of gene expression Multidisciplinary biology Chromosome Biology Drosophila Melanogaster Messenger RNA High-Throughput Nucleotide Sequencing RNA sequencing Animal Models Genomics Spermatids Insects Nucleic acids Meiosis Cell Processes Drosophila Drosophila melanogaster Transcription Initiation Site Cellular Types Drosophila Protein Polypyrimidine Tract-Binding Protein Research Article Arthropoda Molecular Sequence Data Real-Time Polymerase Chain Reaction Research and Analysis Methods 03 medical and health sciences Model Organisms Genetics Animals Polypyrimidine tract-binding protein RNA Messenger Spermatogenesis Molecular Biology Techniques Molecular Biology Binding Sites Base Sequence Gene Expression Profiling lcsh:R Organisms Biology and Life Sciences Cell Biology Reverse Transcriptase-Polymerase Chain Reaction biology.organism_classification Invertebrates Exon skipping Sperm Gene expression profiling 030104 developmental biology Gene Ontology Germ Cells Gene Expression Regulation Animal Genomics Mutation biology.protein RNA lcsh:Q Transcriptome 030217 neurology & neurosurgery

description

The Drosophila polypyrimidine tract-binding protein (dmPTB or hephaestus) plays an important role during spermatogenesis. The heph2 mutation in this gene results in a specific defect in spermatogenesis, causing aberrant spermatid individualization and male sterility. However, the array of molecular defects in the mutant remains uncharacterized. Using an unbiased high throughput sequencing approach, we have identified transcripts that are misregulated in this mutant. Aberrant transcripts show altered expression levels, exon skipping, and alternative 5' ends. We independently verified these findings by reverse-transcription and polymerase chain reaction (RT-PCR) analysis. Our analysis shows misregulation of transcripts that have been connected to spermatogenesis, including components of the actomyosin cytoskeletal apparatus. We show, for example, that the Myosin light chain 1 (Mlc1) transcript is aberrantly spliced. Furthermore, bioinformatics analysis reveals that Mlc1 contains a high affinity binding site(s) for dmPTB and that the site is conserved in many Drosophila species. We discuss that Mlc1 and other components of the actomyosin cytoskeletal apparatus offer important molecular links between the loss of dmPTB function and the observed developmental defect in spermatogenesis. This study provides the first comprehensive list of genes misregulated in vivo in the heph2 mutant in Drosophila and offers insight into the role of dmPTB during spermatogenesis.

year	journal	country	edition	language
2015-11-11	PLoS ONE

10.1371/journal.pone.0150768 http://europepmc.org/articles/PMC4778870?pdf=render