D'Ippolito, Robert AnthonyMinamino, NaokiRivera-Casas, CiroCheema, Manjinder S.Bai, Dina L.Kasinsky, Harold E.Shabanowitz, JeffreyEirin-Lopez, Jose M.Ueda, TakashiHunt, Donald F.Ausio, Juan2021-01-282021-01-2820192019D’Ippolito, R. A., Minamino, N., Rivera-Casas, C., Cheema, M. S., Bai, D. L., Kasinsky, H. E., … Ausió, J. (2019). Protamines from liverwort are produced by post -translational cleavage and C-terminal di-aminopropanelation of several male germ-specific H1 histones. Journal of Biological Chemistry, 294(44), 16364-16373. https://doi.org/10.1074/jbc.RA119.010316.https://doi.org/10.1074/jbc.RA119.010316http://hdl.handle.net/1828/12613Protamines are small, highly-specialized, arginine-rich, and intrinsically-disordered chromosomal proteins that replace histones during spermiogenesis in many organisms. Previous evidence supports the notion that, in the animal kingdom, these proteins have evolved from a primitive replication-independent histone H1 involved in terminal cell differentiation. Nevertheless, a direct connection between the two families of chromatin proteins is missing. Here, we primarily used electron transfer dissociation MS-based analyses, revealing that the protamines in the sperm of the liverwort Marchantia polymorpha result from post-translational cleavage of three precursor H1 histones. Moreover, we show that the mature protamines are further post-translationally modified by di-aminopropanelation, and previous studies have reported that they condense spermatid chromatin through a process consisting of liquid-phase assembly likely involving spinodal decomposition. Taken together, our results reveal that the interesting evolutionary ancestry of protamines begins with histone H1 in both the animal and plant kingdoms.en14-3-3 proteinmass spectrometry (MS)histoneelectron microscopy (EM)chromatographydi-aminopropanelationhistone H1Marchantiamass spectrometryprotaminesArticle