This is supplementary data accompanying a manuscript submission. Recent work on the same topic is described in a pre-print: "The proteomic landscape of centromeric chromatin reveals an essential role for the Ctf19CCAN complex in meiotic kinetochore assembly" Weronika E. Borek, Nadine Vincenten, Eris Duro, Vasso Makrantoni, Christos Spanos, Krishna K. Sarangapani, Flavia de Lima Alves, David A. Kelly, Charles L. Asbury, Juri Rappsilber, Adele L. Marston bioRxiv 2020.06.23.167395; doi: https://doi.org/10.1101/2020.06.23.167395 .

This figshare record contains 4 zipped folders: C terminal constructs Sanger sequencing.zip, Parental vectors Sanger sequencing.zip, Untagged and N terminal constructs Sanger sequencing.zip and Licor Imaging.zip.
The Sanger sequencing folders contain data files in .dna, .ab1 and .seq file formats.The Licor Imaging folder contains data files in .tif, .txt and .jpg file format.The data record also includes 3 files in .docx file format, and 4 tables in .xlsx file format.The document Primary-Data-Index_.docx includes information about what is contained in each folder and data file.The data file names have the following structure: sample name - primer - sequencing name.
Below is a short description of the contents of each of the folders, the .docx files and the .xlsx files:1. Cloning FH and HF containing-vectors.docx: Protocol and lab notes for cloning and screening colonies for the parental tagging vectors2. C terminal tagged and eGFP construct cloning.docx: Protocol and original gel images for cloning and screening colonies for C terminally tagged constructs. Untagged, FH and HF eGFP constructs gel images are also included in this file.3. Untagged and N-terminal tagged construct cloning.docx: Protocol and original gel images for cloning and screening colonies for untagged and N-terminal tagged constructs.4. Parental vectors Sanger sequencing.zip: Original Sanger sequencing files for confirming the parental tagging vectors.5. C terminal constructs Sanger sequencing.zip: Original Sanger sequencing files for confirming C terminally tagged constructs, provided embedded in SnapGene files as well as original files6. Untagged and N terminal constructs Sanger sequencing.zip: Original Sanger sequencing files confirming untagged and N-terminally tagged constructs.7. Licor imaging.zip: Original scans of the western blots performed.Table 3.xlx includes data on the quantification of protein abundance, following different induction time courses.Table 4.xlsx includes the mass spectrometry data for the expression of proteins tagged at the N-terminus. Table 5.xlsx includes the oligonucleotide sequences used in this study.Table 6.xlsx includes the sequences of fusion protein open reading frames (ORFs).
Study aims and methodology: SARS-CoV-2 is a large positive-sense, single-stranded RNA virus that encodes four structural proteins, several accessory proteins, and sixteen nonstructural proteins (nsp1-16). The latter are mainly engaged in enzymatic activities important for translation and replication of the RNA genome. In addition, nonstructural proteins also target host RNA metabolism in order to manipulate cellular gene expression or facilitate immune evasion. Understanding these pathways in greater detail will be an important step in the development of antiviral therapies.In this study, the authors generated a series of synthetic, codon-optimized constructs for 14 different viral proteins that are expected to interact with RNA or RNA binding proteins. To remove the need for error-prone PCR steps, they devised a cloning scheme in which a single synthetic construct could be used to generate untagged, N-, or C-terminally tagged versions of the protein. Using these vectors, the authors generated and tested a series of human cell lines with the viral open reading frames (ORFs) stably integrated, under the control of an inducible Tet-On promoter.For details on the methodology, please read the related article.
Software needed to access data: Datasets in .dna and .ab1 file format can be accessed using the SnapGene software.

This figshare record contains 4 zipped folders: C terminal constructs Sanger sequencing.zip, Parental vectors Sanger sequencing.zip, Untagged and N terminal constructs Sanger sequencing.zip and Licor Imaging.zip.
The Sanger sequencing folders contain data files in .dna, .ab1 and .seq file formats.The Licor Imaging folder contains data files in .tif, .txt and .jpg file format.The data record also includes 3 files in .docx file format, and 4 tables in .xlsx file format.The document Primary-Data-Index.docx includes information about what is contained in each folder and data file.The data file names have the following structure: sample name - primer - sequencing name.
Below is a short description of the contents of each of the folders, the .docx files and the .xlsx files:1. Cloning FH and HF containing-vectors.docx: Protocol and lab notes for cloning and screening colonies for the parental tagging vectors2. C terminal tagged and eGFP construct cloning.docx: Protocol and original gel images for cloning and screening colonies for C terminally tagged constructs. Untagged, FH and HF eGFP constructs gel images are also included in this file.3. Untagged and N-terminal tagged construct cloning.docx: Protocol and original gel images for cloning and screening colonies for C terminally tagged constructs.4. Parental vectors Sanger sequencing.zip: Original Sanger sequencing files for confirming the parental tagging vectors.5. C terminal constructs Sanger sequencing.zip: Original Sanger sequencing files for confirming C terminally tagged constructs, provided embedded in SnapGene files as well as original files6. Untagged and N terminal constructs Sanger sequencing.zip: Original Sanger sequencing files confirming untagged and N-terminally tagged constructs.7. Licor imaging.zip: Original scans of the western blots performed.Table 3.xlx includes data on the quantification of protein abundance, following different induction time courses.Table 4.xlsx includes the mass spectrometry data for the expression of proteins tagged at the N-terminus. Table 5.xlsx includes the oligonucleotide sequences used in this study.Table 6.xlsx includes the sequences of fusion protein open reading frames (ORFs).
Study aims and methodology: SARS-CoV-2 is a large positive-sense, single-stranded RNA virus that encodes four structural proteins, several accessory proteins, and sixteen nonstructural proteins (nsp1-16). The latter are mainly engaged in enzymatic activities important for translation and replication of the RNA genome. In addition, nonstructural proteins also target host RNA metabolism in order to manipulate cellular gene expression or facilitate immune evasion. Understanding these pathways in greater detail will be an important step in the development of antiviral therapies.In this study, the authors generated a series of synthetic, codon-optimized constructs for 14 different viral proteins that are expected to interact with RNA or RNA binding proteins. To remove the need for error-prone PCR steps, they devised a cloning scheme in which a single synthetic construct could be used to generate untagged, N-, or C-terminally tagged versions of the protein. Using these vectors, the authors generated and tested a series of human cell lines with the viral open reading frames (ORFs) stably integrated, under the control of an inducible Tet-On promoter.For details on the methodology, please read the related article.
Software needed to access data: Datasets in .dna and .ab1 file format can be accessed using the SnapGene software.

This figshare record contains 4 zipped folders: C terminal constructs Sanger sequencing.zip, Parental vectors Sanger sequencing.zip, Untagged and N terminal constructs Sanger sequencing.zip and Licor Imaging.zip.
The Sanger sequencing folders contain data files in .dna, .ab1 and .seq file formats.The Licor Imaging folder contains data files in .tif, .txt and .jpg file format.The data record also includes 3 files in .docx file format, and 4 tables in .xlsx file format.The document Primary-Data-Index_.docx includes information about what is contained in each folder and data file.The data file names have the following structure: sample name - primer - sequencing name.
Below is a short description of the contents of each of the folders, the .docx files and the .xlsx files:1. Cloning FH and HF containing-vectors.docx: Protocol and lab notes for cloning and screening colonies for the parental tagging vectors2. C terminal tagged and eGFP construct cloning.docx: Protocol and original gel images for cloning and screening colonies for C terminally tagged constructs. Untagged, FH and HF eGFP constructs gel images are also included in this file.3. Untagged and N-terminal tagged construct cloning.docx: Protocol and original gel images for cloning and screening colonies for untagged and N-terminal tagged constructs.4. Parental vectors Sanger sequencing.zip: Original Sanger sequencing files for confirming the parental tagging vectors.5. C terminal constructs Sanger sequencing.zip: Original Sanger sequencing files for confirming C terminally tagged constructs, provided embedded in SnapGene files as well as original files6. Untagged and N terminal constructs Sanger sequencing.zip: Original Sanger sequencing files confirming untagged and N-terminally tagged constructs.7. Licor imaging.zip: Original scans of the western blots performed.Table 3.xlx includes data on the quantification of protein abundance, following different induction time courses.Table 4.xlsx includes the mass spectrometry data for the expression of proteins tagged at the N-terminus. Table 5.xlsx includes the oligonucleotide sequences used in this study.Table 6.xlsx includes the sequences of fusion protein open reading frames (ORFs).
Study aims and methodology: SARS-CoV-2 is a large positive-sense, single-stranded RNA virus that encodes four structural proteins, several accessory proteins, and sixteen nonstructural proteins (nsp1-16). The latter are mainly engaged in enzymatic activities important for translation and replication of the RNA genome. In addition, nonstructural proteins also target host RNA metabolism in order to manipulate cellular gene expression or facilitate immune evasion. Understanding these pathways in greater detail will be an important step in the development of antiviral therapies.In this study, the authors generated a series of synthetic, codon-optimized constructs for 14 different viral proteins that are expected to interact with RNA or RNA binding proteins. To remove the need for error-prone PCR steps, they devised a cloning scheme in which a single synthetic construct could be used to generate untagged, N-, or C-terminally tagged versions of the protein. Using these vectors, the authors generated and tested a series of human cell lines with the viral open reading frames (ORFs) stably integrated, under the control of an inducible Tet-On promoter.For details on the methodology, please read the related article.
Software needed to access data: Datasets in .dna and .ab1 file format can be accessed using the SnapGene software.

Data in support of manuscript: "Lamin A molecular compression and sliding as mechanisms behind nucleoskeleton elasticity" Lamin A is a nuclear intermediate filament protein critical for nuclear architecture and mechanics and mutated in a wide range of human diseases. Yet little is known about the molecular architecture of lamins and mechanisms of their assembly. Here we use cross-linking SILAC mass spectrometry to determine interactions within lamin dimers and between dimers in higher-order polymers. We find evidence for a compression mechanism where coiled coils in the lamin A rod can slide onto each other to contract rod length, likely driven by a wide range of electrostatic interactions with the flexible linkers between coiled coils. Similar interactions occur with unstructured regions flanking the rod domain during oligomeric assembly. Mutations linked to human disease block these interactions, suggesting that this spring-like contraction can explain in part the dynamic mechanical stretch and flexibility properties of the lamin polymer and other intermediate filament networks.

Supplementary data showing protein names, SILAC ratios, peptide counts, and peptide intensities identified by SILAC MS in T7-TRIM25, T7-TRIM25Î RBD, and T7-TRIM25â +â RNases co-immunoprecipitation assays. (XLSX 22 kb)

Supplementary data showing protein names, SILAC ratios, peptide counts, and peptide intensities identified by SILAC MS in T7-TRIM25, T7-TRIM25Î RBD, and T7-TRIM25â +â RNases co-immunoprecipitation assays. (XLSX 22 kb)

Supplementary data showing protein names, SILAC ratios (more than two-fold), peptide counts, and peptide intensities identified by SILAC MS in wild-type and TRIM25 KO HeLa cells. (XLSX 33 kb)

Supplementary data showing gene names and number of RNA reads derived from three independent T7-TRIM25 CLIP-seq experiments. (XLSX 117 kb)

Supplementary data showing gene names and number of RNA reads derived from three independent T7-TRIM25 CLIP-seq experiments. (XLSX 117 kb)