Data from: Caspar specifies primordial germ cell count and identity in Drosophila melanogaster
Data files
Feb 05, 2025 version files 895.06 MB
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Imaging_Data.zip
885.96 MB
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Mass_Spec_Data.zip
8.90 MB
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README.md
2.91 KB
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Sequencing_Data_(Caspar).zip
197.28 KB
Abstract
Repurposing of pleiotropic factors during execution of diverse cellular processes has emerged as a regulatory paradigm. Embryonic development in metazoans is controlled by maternal factors deposited in the egg during oogenesis. Here, we explore maternal role(s) of Caspar (Casp), the Drosophila orthologue of human Fas-associated factor-1 (FAF1) originally implicated in host-defense as a negative regulator of NF-κB signaling. Maternal loss of either Casp or it’s protein partner, Transitional endoplasmic reticulum 94 (TER94) leads to partial embryonic lethality correlated with aberrant centrosome behavior, cytoskeletal abnormalities, and defective gastrulation. Although ubiquitously distributed, both proteins are enriched in the primordial germ cells (PGCs), and in keeping with the centrosome problems, mutant embryos display a significant reduction in the PGC count. Moreover, the total number of pole buds is directly proportional to the level of Casp. Consistently, it’s ‘loss’ and ‘gain’ results in respective reduction and increase in the Oskar protein levels, the master determinant of PGC fate. To elucidate this regulatory loop, we analyzed several known components of mid-blastula transition and identify the translational repressor Smaug, a zygotic regulator of germ cell specification, as a potential critical target. We present a detailed structure-function analysis of Casp aimed at understanding its novel involvement during PGC development.
README: Supplementary Data
https://doi.org/10.5061/dryad.zs7h44jkf
Description of the data and file structure
The Uploaded dataset contains three sets of data - as individual zipped folders:
- Imaging File folder (Each Figure in the Manuscript is in a labelled folder in the data set).
- Sequencing Data folder (For Caspar pUASp clones)
- Mass Spec file folder (for Caspar & Mock IPs).
The Manuscript is linked here (https://elifesciences.org/reviewed-preprints/98584), and the experimental methodology is described in Materials & Methods.
Files and variables
Three folders; Imaging Data.zip, Mass spec data.zip, Sequencing Data (caspar).zip
Description: The Uploaded data contains three datasets (folders).
- Folder 'Imaging Data'. The folder contain Microscopy Imaging Files (Each Figure in the Manuscript is in a labelled folder). The Subfolders are labelled as FIG 1, FIG 2, FIG 3, FIG4, FIG 5, FIG 6, FIG 8, FIG 2, FIG S3, FIG S4. Each subfolder either has raw imaging data or additional subfolders that relate to specific Figures. For example FIG 6 has three sub-folders FIG 6A, 6B & 6C. Each subfolder has an imaging file in .lsm (Leica) format. Leica format files can be opened by ImageJ/Fiji.
- Folder 'Sequencing Data (Caspar constructs)'. The names of the sequencing files correspond to the experimental constructs used in the manuscript. Each sequencing file is in FASTA (.fsta) format. One .docx file (pUASp Caspar constructs) has the construct names and the sequence of the fusion protein expressed from the starting ATG to terminating TGA codons. All constructs are in pUASp vector backbone. I did not include the backbone sequence in the column2. The second .docx file (pUASp casp sequencing results) contains the names and sequences of the sequencing files.
- Folder 'Mass Spec Data': The folder contains two files, CaspFDR.xlsx and IgGFDR.xlsx. The first is the experimental Immunoprecipitate(IP) using the Caspar antibody while the second file contains the control IP using pre-immune serum. The files have been generated by Sciex Protein Pilot 2.0. The files contain the following Tabs: Protein FDR Summary, Distinct Protein FDR Summary, Spectrum FDR Summary, Protein Summary, Proteins, Peptide Summary, Distinct Peptide Summary, Feature Summary, Single Column Summary. The IgGFDR (mock IP) file, has a custom, used created 'Sorted' Tab with proteins sorted by number of peptides counted while the Casp_FDR has a similar 'sorted' and a 'IgG vs Casp' tab. The 'IgG vs Casp' tab lists, in yellow highlight the unique proteins discovered, after igG correction.
Code/software
None
Access information
Other publicly accessible locations of the data:
- Not Applicable
Data was derived from the following sources:
- Not Applicable