DOI: 10.5061/dryad.8sf7m0csb
title: RNA sequencing data from the guts of Drosophila wildtype and CG3740/nazo double mutant 20-day-old males

description: Lipid dyshomeostasis has been implicated in a variety of diseases ranging
 from obesity to neurodegenerative disorders such as Neurodegeneration with
 Brain Iron Accumulation (NBIA). Here, we uncover the physiological role of
 Nazo, the Drosophilamelanogaster homolog of the NBIA-mutated protein –
 c19orf12, whose function has been elusive. Ablation of Drosophila c19orf12
 homologs leads to dysregulation of multiple lipid metabolism genes. nazo
 mutants exhibit markedly reduced gut lipid droplet and whole-body
 triglyceride contents. Consequently, they are sensitive to starvation and
 oxidative stress. Nazo is required for maintaining normal levels of
 Perilipin-2, an inhibitor of the lipase – Brummer. Concurrent knockdown of
 Brummer or overexpression of Perilipin-2 rescues the nazo phenotype,
 suggesting that this defect, at least in part, may arise from diminished
 Perilipin-2 on lipid droplets leading to aberrant Brummer-mediated
 lipolysis. Our findings potentially provide novel insights into the role
 of c19orf12 as a possible link between lipid dyshomeostasis and
 neurodegeneration, particularly in the context of NBIA.
description: The samples( guts) were dissected in ice-cold PBS and
 flash-frozen in liquid nitrogen followed by RNA extraction using a
 standard RNA extraction kit (Qiagen) before being processed. 
description: # Title: RNA Sequencing data from the guts of Drosophila wildtype and
 CG3740/nazo double mutant 20 day old males. * Authors: Sreejith
 Perinthottathil, Rajnish Bharadwaj * Date created: 2022-09-19 * Date
 modified: 2022-09-20 \[Access this dataset on Dryad](Dataset DOI link)
 --- * Dataset Title: Data
 for the article "Nazo, the Drosophila homolog of the NBIA-mutated
 protein – c19orf12, is required for triglyceride homeostasis" *
 Persistent Identifier: *
 Dataset Contributors: * Creators: Perinthottathil Sreejith, Sara Lolo,
 Kristen R. Patten, Maduka Gunasinghe, Neya More, Leo J.Pallanck, Rajnish
 Bharadwaj * Date of Issue: 2022-09-30 * Publisher: University of Rochester
 ## Contact Information * Name: Rajnish Bharadwaj * Affiliations: Pathology
 and Lab Medicine, University of Rochester Medical Center * ORCID
 ID: * Email:
 * Alternative Contact:
 postdoctoral PI * Name: Sreejith Perinthottathil * Affiliations: Pathology
 and Lab Medicine, University of Rochester Medical Center * ORCID ID:
 * Email:
 # Additional Dataset Metadata ##
 Acknowledgements * Funding sources: This research was partly funded by
 Neurodegeneration with Brain Iron Disorders Association grant. ## Dates
 and Locations * Dates of data collection: The samples were collected on
 Jan 2020 in Rochester NY * Geographic locations of data collection: Wet
 lab work performed at University of Rochester Medical Center. Samples were
 sequenced at the Genomic Resource Center, University of Rochester Medical
 Center. --- # Methodological Information * Methods of data
 collection/generation: see manuscript for details Manuscript in Biorxiv:
 Nazo, the Drosophila
 homolog of the NBIA-mutated protein – c19orf12, is required for
 triglyceride homeostasis Perinthottathil Sreejith, Sara Lolo, Kristen R.
 Patten, Maduka Gunasinghe, Neya More, Leo J. Pallanck, Rajnish Bharadwaj
 bioRxiv 2022.09.29.510106; doi:
 The data represents the
 RNA sequencing results of 20-day-old male guts dissected in PBS followed
 by the RNA sequencing analysis as described in the manuscript. Briefly,
 Total RNA was isolated from triplicates of 20 freshly dissected guts of
 20-day-old WT and CG3740/nazo mutant males using the RNeasy Plus Mini Kit
 (Qiagen, Valencia, CA) per manufacturer’s recommendations. RNA-seq
 analysis was performed at UR Genomics Research Center. The total RNA
 concentration was determined with the NanopDrop 1000 spectrophotometer
 (NanoDrop, Wilmington, DE) and RNA quality was assessed with the Agilent
 Bioanalyzer (Agilent, Santa Clara, CA). The TruSeq Stranded mRNA Sample
 Preparation Kit (Illumina, San Diego, CA) was used for next generation
 sequencing library construction per manufacturer’s protocols. Briefly,
 mRNA was purified from 200ng total RNA with oligo-dT magnetic beads and
 fragmented. First-strand cDNA synthesis was performed with random hexamer
 priming followed by second-strand cDNA synthesis using dUTP incorporation
 for strand marking. End repair and 3` adenylation was then performed on
 the double stranded cDNA. Illumina adaptors were ligated to both ends of
 the cDNA, purified by gel electrophoresis and amplified with PCR primers
 specific to the adaptor sequences to generate amplified libraries of
 approximately 200-500bp in size. The amplified libraries were hybridized
 to the Illumina flow cell and single end reads of 75nt were generated for
 each sample using Illumina’s NextSeq550 sequencer. ## Description of the
 data and file structure File Structure: RB_SP RNAseq&gt; subfolder
 1&gt;deliv_Bharadwaj_021720_results&gt; Sub folder&gt; deSeq2&gt; Sub
 Folder&gt; Star_featureCounts&gt; Sub folder -contains 5 files RB_SP
 RNAseq&gt; subfolder 2&gt;deliv_Bharadwaj_021720_raw.tar &gt;
 Subfolder&gt; deliv_Bharadwaj_021720_raw&gt; sub folder cp\ontains 6 files
 Describe relationships between data files, missing data codes, other
 abbreviations used. Be as descriptive as possible. Abbreviations: * RBmut=
 Double Mutants * RBWT = Wildtype * dn= down regulated genes * up=
 upregulated genes * NormCounts = Normalized counts * volcano = Volcano
 plot File names in folder RB_SP RNAseq&gt; subfolder
 1&gt;deliv_Bharadwaj_021720_results&gt; Sub folder&gt; deSeq2&gt; Sub
 Folder&gt; Star_featureCounts 1. deSeq2_counts Excel file 2.
 deSeq2_NormCounts Excel file 3. deSeq2_RBMut_vs_RBWT Excel file 4.
 deSeq2_RBMut_vs_RBWT_heatmap PDF file 5. deSeq2_rlog_NormCounts Excel file
 FIle names in folder RB_SP RNAseq&gt; subfolder
 2&gt;deliv_Bharadwaj_021720_raw.tar &gt; Subfolder&gt;
 deliv_Bharadwaj_021720_raw&gt; 1. RBmut1_R1.fastq 2. RBmut2_R1.fastq 3.
 RBmut3_R1.fastq 4. RBWT1_R1.fastq 5. RBWT2_R1.fastq 6. RBWT3_R1.fastq
 Brief description of the above files: Each folder is a FASTQ sequence
 trace of each replicate of either Wiltype(WT) guts or Double mutant (mut)
 guts. ## Sharing/Access information Description of the datasets ####
 deSeq2_counts Read counts for the sum of reads associated with each of the
 exons (feature) that “belong” to that gene detected in the RNA seq of guts
 in triplicate in Wildtype(WT) and Double mutant(RBmut). ####
 deSeq2_NormCounts Normalized counts for the number of species of genes
 detected in the RNA seq of guts in triplicates in Wildtype(WT) and Double
 mutants (RBmut) It gives the number of reads that align to a feature after
 correcting for sequencing depth and transcriptome composition bias. N ####
 deSeq2_RBMut_vs_RBWT Comparison of the expression of genes expressed in
 the Wildtype and Double mutant with the basemean values-the average of the
 normalized count values for all samples, log fold change- The change in
 expression of the gene, pvalues-a measure of how likely it is to get the
 data if no real difference existed and adjusted pvalues-The p-value
 adjusted (padj) column contains the p-values, adjusted for multiple
 testing with the Benjamini-Hochberg procedure. ####
 deSeq2_RBMut_vs_RBWT_heatmap Heat map of the comparison of the expression
 of Wildtype(RBWT) and Double mutant(RBmut) #### deSeq2_rlog_NormCounts
 Regularized log values of the genes expressed in the Wiltype and Double
 mutant guts.