MassIVEapplication/xmlftp://massive-ftp.ucsd.edu/v04/MSV000088458/primary200OK1000ProteomicsJim PesaventoOrbitrap Fusion LumosChlamydomonas Reinhardtii (ncbitaxon:3055)https://massive.ucsd.edu/ProteoSAFe/dataset.jsp?task=ebda5a2a6da4462c823f5625ea6be6f0Saint Mary's College of Californiajjp6@stmarys-ca.eduMassIVE6UNIMOD:37 - "Tri-Methylation."UNIMOD:34 - "Methylation."UNIMOD:36 - "Di-Methylation."UNIMOD:1 - "Acetylation."UNIMOD:121 - "Ubiquitinylation residue."We present an updated analysis of the linker and core histone proteins and their proteoforms in the green microalga <i>Chlamydomonas reinhardtii</i> by top-down mass spectrometry (TDMS). The combination of high-resolution liquid chromatographic separation, robust fragmentation, high mass spectral resolution, the application of a custom search algorithm, and extensive manual analysis enabled the characterization of 86 proteoforms across all four core histones H2A, H2B, H3, and H4 and the linker histone H1. All canonical H2A paralogs, which vary in their C-termini, were identified, along with the previously unreported noncanonical variant H2A.Z that had high levels of acetylation and C-terminal truncations. Similarly, a majority of the canonical H2B paralogs were identified, along with a smaller noncanonical variant, H2B.v1, that was highly acetylated. Histone H4 exhibited a novel acetylation profile that differs significantly from that found in other organisms. A majority of H3 was monomethylated at K4 with low levels of co-occuring acetylation, while a small fraction of H3 was trimethylated at K4 with high levels of co-occuring acetylation.The heterodimeric tumor-suppressor complex BRCA1/BARD1 exhibits E3 ubiquitin ligase activity and participates in cell proliferation and chromosome stability control by incompletely defined mechanisms. Here we show that, in both mammalian cells and Xenopus egg extracts, BRCA1/BARD1 is required for mitotic spindle-pole assembly and for accumulation of TPX2, a major spindle organizer and Ran target, on spindle poles. This function is centrosome independent, operates downstream of Ran GTPase, and depends upon BRCA1/BARD1 E3 ubiquitin ligase activity. Xenopus BRCA1/BARD1 forms endogenous complexes with three spindle-pole proteins, TPX2, NuMA, and XRHAMM--a known TPX2 partner--and specifically attenuates XRHAMM function. These observations reveal a previously unrecognized function of BRCA1/BARD1 in mitotic spindle assembly that likely contributes to its role in chromosome stability control and tumor suppression.An Improved Top-Down Mass Spectrometry Characterization of <i>Chlamydomonas reinhardtii</i> Histones and Their Post-translational Modifications.The BRCA1/BARD1 heterodimer modulates ran-dependent mitotic spindle assembly.Rommelfanger Sarah R SR, Zhou Mowei M, Shaghasi Henna H, Tzeng Shin-Cheng SC, Evans Bradley S BS, Paša-Tolić Ljiljana L, Umen James G JG, Pesavento James J JJJoukov Vladimir V, Groen Aaron C AC, Prokhorova Tatyana T, Gerson Ruth R, White Erinn E, Rodriguez Alison A, Walter Johannes C JC, Livingston David M DMliquid chromatography tandem mass spectroscopy, LC-MS2, rock salt, determination, sales, acid, LC-MS/MS, Histone, LC-MS-MS, Histone H2b, Histone H2a, common salt, LC/MS/MS, halite, Natriumchlorid, ionic compounds, LC-MSMS, chemical analysis, Chlamydomona, Msal-1, table salt, NaCl, Histone H3.3, Sal, Acid, LCMSMS, Spalt, ionic compound, Saeuren, acide, Salt, chlorure de sodium, acids, Msal, B130022O04Rik, Papers., Salze, acido, natrii chloridum, sel, Salz, Histone H1(s), salts, sal, cloruro sodico, salt, Dietary Sodium, liquid chromatography-tandem mass spectroscopy, sels, Kochsalz, Histone H5, Histone H4, Histone H7, liquid chromatography tandem mass spectrometry, assay, Saeure, Histone H1, Histone H3XH2AZ, HISTONE H2B, determination, H2A/z, Chlamydomonas, Hist2av, Gene, reinhardtii, gamma-His2Av, Spectrum Analyses, H2A.Z-1, His2, Histone H2b, H1(any), Histone H2a, H2A|O, reduced, Gene Products, Mass, CG5499, Low, Analysis, tiny, h2afx, Separated, Histone H3.3, Mass Spectroscopy, HIS2AV, Mass Spectrum Analysis, H2A|q, reinhardius, Divorced, me75, H2A.F/Z, H2AV_DROM, gamma-HIS2AV, Analyses, MRN17_11, HisH2Av, hypoplasia, Acetylations, h2AvD, HIS2AVD, H2B.1, Divorces, H2A|z, D17Mit170, T1, Hist, MRN17.11, F8N16.2, l(3)05146, Chlamydomonas reihhardtii, H2a.z-1, H2AvD, H2BFQ, Chlamydomonas reinhardii, Histone H5, Histone H4, Histone H7, HIS, His, liquid, Chlamydomonas smithii, Histone H1, Histone H3, DmelCG5499, H2AX, small, H2AZ, cou, HIST2H2AA, l(3)97Dd, H2avD, histone H4, Proteins, acetylation, F8N16_2, his, H2A.2, H2A, H2BQ, H2B, Chlamydomonas reinhardtius, Tl3, Tl2, Histone, Spectrum Analysis, H2Av, Spectroscopy, MS, gamma-H2Av, Lr, H2AFQ, Algorithm, chemical analysis, Protein, H2AFO, H2BGL105, core, H2A-GL101, H2a, H2b, H2a-615, Mass Spectrum Analyses, gammaH2Av, H2AV, H2av, l(3)810, Mass Spectrum, Separation, l(3)L1602, underdeveloped, histone B2, H2A.Z, Separations, H2A.X, H2a.V, Spectrometry, h2a, His2AvD, Chlamydomonas reinhardius, H2A/O, Histone H1(s), Protein Gene Products, h2a.zl1, h2a.zl2, Gene Proteins, H2Az, 5499, His2AV, H2A.z, Bra, assay, H2A/q, Acetylations., Chlamydomonas smithii R.W.Howshaw & H.Ettl, GL105, Chlamydomonas reinhardtii P.A.Dangeard, HISTONE 2BPost Translational Amino Acid Modification, Post-Translational Modification, Post-Translational Protein Modifications, Chlamydomonas, Modifications, Post-Translational Protein Modification, reinhardtii, Spectrum Analyses, Chlamydomonas reinhardtius, Spectrum Analysis, Histone, Protein Processing, Posttranslational, Spectroscopy, Histone H2b, Histone H2a, MS, Post Translational Modifications, Posttranslational Modification., Mass, Post Translational Modification, Post-Translational Modifications, Analysis, Protein Modification, Mass Spectrum Analyses, Mass Spectroscopy, Histone H3.3, Post Translational, Mass Spectrum Analysis, Modification, Post-Translational Protein, Mass Spectrum, reinhardius, Analyses, Post-Translational, Post Translational Protein Processing, Processing, Spectrometry, Chlamydomonas reinhardius, Posttranslational Protein Processing, Histone H1(s), Protein Modifications, Post-Translational Amino Acid Modification, Chlamydomonas reihhardtii, Post Translational Protein Modification, Amino Acid Modification, Posttranslational Amino Acid Modification, Chlamydomonas reinhardii, Post-Translational Protein Processing, Histone H5, Histone H4, Posttranslational Protein, Histone H7, Posttranslational Modifications, Chlamydomonas smithii R.W.Howshaw & H.Ettl, Chlamydomonas reinhardtii P.A.Dangeard, Chlamydomonas smithii, Histone H1, Histone H3Protein Gene Products, Gene Proteins, Histone H2b, Histone H2a, determination, Chlamydomona., chemical analysis, Protein, Histone H5, Gene Products, Proteins, Histone H4, Histone H7, Gene, assay, Histone H1(s), Histone H1, Histone, Histone H3.3, Histone H31falseTop-down analysis of histone proteins from asynchronous chlamydomonasThese are salt-and-acid histone extracts from asynchronously grown Chlamydomonas measured by LC-MS/MS via top-down analysis. These 5 runs are biological repeats used in our JASMS 2021 paper (Rommelfanger et al.).Fri Nov 26 13:37:00 GMT 2021MSV00008845834165968