Discovery of Dehydroamino Acid Residues in the Capsid and Matrix Structural Proteins of HIV-1.

Human immunodeficiency virus type 1 (HIV-1) remains a deadly infectious disease despite existing antiretroviral therapies. A comprehensive understanding of the specific mechanisms of viral infectivity remains elusive and currently limits the development of new and effective therapies. Through in-depth proteomic analysis of HIV-1 virions, we discovered the novel post-translational modification of highly conserved residues within the viral matrix and capsid proteins to the dehydroamino acids, dehydroalanine and dehydrobutyrine. We further confirmed their presence by labeling the reactive alkene, characteristic of dehydroamino acids, with glutathione via Michael addition. Dehydroamino acids are rare, understudied, and have been observed mainly in select bacterial and fungal species. Until now, they have not been observed in HIV proteins. We hypothesize that these residues are important in viral particle maturation and could provide valuable insight into HIV infectivity mechanisms.

[1]  K. P. Chooi,et al.  LanCLs add glutathione to dehydroamino acids generated at phosphorylated sites in the proteome , 2021, Cell.

[2]  Rebecca A. Scheck,et al.  A chemical probe for dehydrobutyrine. , 2020, Angewandte Chemie.

[3]  M. Fujino,et al.  HIV-1 matrix mutations that alter gag membrane binding modulate mature core formation and post-entry events. , 2019, Virology.

[4]  K. Schey,et al.  Quantification of thioether‐linked glutathione modifications in human lens proteins , 2018, Experimental eye research.

[5]  Michael R Shortreed,et al.  Enhanced Global Post-translational Modification Discovery with MetaMorpheus. , 2018, Journal of proteome research.

[6]  Brian L. Frey,et al.  Global Post-Translational Modification Discovery , 2016, Journal of proteome research.

[7]  P. Ciborowski,et al.  Proteomics, biomarkers, and HIV‐1: A current perspective , 2015, Proteomics. Clinical applications.

[8]  S. Silverman,et al.  Phosphoserine Lyase Deoxyribozymes: DNA-Catalyzed Formation of Dehydroalanine Residues in Peptides. , 2015, Journal of the American Chemical Society.

[9]  T. Hope,et al.  HIV-1 capsid: the multifaceted key player in HIV-1 infection , 2015, Nature Reviews Microbiology.

[10]  Ming Li.  Proteomics in the investigation of HIV‐1 interactions with host proteins , 2015, Proteomics. Clinical applications.

[11]  Reinhard Schneider,et al.  The HIV Mutation Browser: A Resource for Human Immunodeficiency Virus Mutagenesis and Polymorphism Data , 2014, PLoS Comput. Biol..

[12]  D. Siodłak.  α,β-Dehydroamino acids in naturally occurring peptides , 2014, Amino Acids.

[13]  T. Nakano,et al.  Phosphorylation of human immunodeficiency virus type 1 capsid protein at serine 16, required for peptidyl-prolyl isomerase-dependent uncoating, is mediated by virion-incorporated extracellular signal-regulated kinase 2. , 2014, The Journal of general virology.

[14]  K. Schey,et al.  Human protein aging: modification and crosslinking through dehydroalanine and dehydrobutyrine intermediates , 2013, Aging cell.

[15]  Robert J. Gifford,et al.  Extreme Genetic Fragility of the HIV-1 Capsid , 2013, Retrovirology.

[16]  S. Gerstberger,et al.  Methods for converting cysteine to dehydroalanine on peptides and proteins , 2011 .

[17]  Mark Yeager,et al.  Atomic level modeling of the HIV capsid , 2010, Nature.

[18]  Alasdair C. Steven,et al.  Visualization of a Missing Link in Retrovirus Capsid Assembly , 2009, Nature.

[19]  D. Bar-Or,et al.  Dehydroalanine derived from cysteine is a common post-translational modification in human serum albumin. , 2008, Rapid communications in mass spectrometry : RCM.

[20]  A. Bukrinskaya.  HIV-1 matrix protein: a mysterious regulator of the viral life cycle. , 2007, Virus research.

[21]  J. Szostak,et al.  Ribosomal synthesis of dehydroalanine-containing peptides. , 2006, Journal of the American Chemical Society.

[22]  David H Perlman,et al.  Reverse transcription-associated dephosphorylation of hepadnavirus nucleocapsids. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[23]  S. Scarlata,et al.  The cysteine residues of HIV-1 capsid regulate oligomerization and cyclophilin A-induced changes. , 2005, Biophysical journal.

[24]  Marc C. Johnson,et al.  The stoichiometry of Gag protein in HIV-1 , 2004, Nature Structural &Molecular Biology.

[25]  Wesley I. Sundquist,et al.  Image reconstructions of helical assemblies of the HIV-1 CA protein , 2022 .

[26]  W. Sundquist,et al.  Assembly and analysis of conical models for the HIV-1 core. , 1999, Science.

[27]  W. Sundquist,et al.  Proteolytic refolding of the HIV‐1 capsid protein amino‐terminus facilitates viral core assembly , 1998, The EMBO journal.

[28]  A. Kingsman,et al.  Structure-function studies of the human immunodeficiency virus type 1 matrix protein, p17 , 1997, Journal of virology.

[29]  V. Vogt,et al.  Analysis of Rous sarcoma virus Gag protein by mass spectrometry indicates trimming by host exopeptidase , 1996, Journal of virology.

[30]  W. Sundquist,et al.  Three-dimensional structure of the human immunodeficiency virus type 1 matrix protein. , 1994, Journal of molecular biology.

[31]  E. Freed,et al.  Single amino acid changes in the human immunodeficiency virus type 1 matrix protein block virus particle production , 1994, Journal of virology.

[32]  C. Anderson,et al.  The human DNA-activated protein kinase phosphorylates simian virus 40 T antigen at amino- and carboxy-terminal sites , 1991, Journal of virology.