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N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB

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Abstract
Protein N-terminal acetylation (Nt-acetylation) is an important mediator of protein function, stability, sorting, and localization. Although the responsible enzymes are thought to be fairly well characterized, the lack of identified in vivo substrates, the occurrence of Nt-acetylation substrates displaying yet uncharacterized N-terminal acetyltransferase (NAT) specificities, and emerging evidence of posttranslational Nt-acetylation, necessitate the use of genetic models and quantitative proteomics. NatB, which targets Met-Glu-, Met-Asp-, and Met-Asn-starting protein N termini, is presumed to Nt-acetylate 15% of all yeast and 18% of all human proteins. We here report on the evolutionary traits of NatB from yeast to human and demonstrate that ectopically expressed hNatB in a yNatB-Delta yeast strain partially complements the natB-Delta phenotypes and partially restores the yNatB Nt-acetylome. Overall, combining quantitative N-terminomics with yeast studies and knockdown of hNatB in human cell lines, led to the unambiguous identification of 180 human and 110 yeast NatB substrates. Interestingly, these substrates included Met-Gln- N-termini, which are thus now classified as in vivo NatB substrates. We also demonstrate the requirement of hNatB activity for maintaining the structure and function of actomyosin fibers and for proper cellular migration. In addition, expression of tropomyosin-1 restored the altered focal adhesions and cellular migration defects observed in hNatB-depleted HeLa cells, indicative for the conserved link between NatB, tropomyosin, and actin cable function from yeast to human.
Keywords
ACTIN, IDENTIFICATION, TROPOMYOSIN, ACETYLATION, YEAST, CELL-MIGRATION, PROTEIN TERMINI, SACCHAROMYCES-CEREVISIAE, METHIONINE AMINOPEPTIDASE, ALPHA-ACETYLTRANSFERASE

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Chicago
Van Damme, Petra, Marta Lasa, Bogdan Polevoda, Cristina Gazquez, Alberto Elosegui-Artola, Duk Soo Kim, Elena De Juan-Pardo, et al. 2012. “N-terminal Acetylome Analyses and Functional Insights of the N-terminal Acetyltransferase NatB.” Proceedings of the National Academy of Sciences of the United States of America 109 (31): 12449–12454.
APA
Van Damme, Petra, Lasa, M., Polevoda, B., Gazquez, C., Elosegui-Artola, A., Kim, D. S., De Juan-Pardo, E., et al. (2012). N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 109(31), 12449–12454.
Vancouver
1.
Van Damme P, Lasa M, Polevoda B, Gazquez C, Elosegui-Artola A, Kim DS, et al. N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA. 2012;109(31):12449–54.
MLA
Van Damme, Petra, Marta Lasa, Bogdan Polevoda, et al. “N-terminal Acetylome Analyses and Functional Insights of the N-terminal Acetyltransferase NatB.” PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 109.31 (2012): 12449–12454. Print.
@article{3048722,
  abstract     = {Protein N-terminal acetylation (Nt-acetylation) is an important mediator of protein function, stability, sorting, and localization. Although the responsible enzymes are thought to be fairly well characterized, the lack of identified in vivo substrates, the occurrence of Nt-acetylation substrates displaying yet uncharacterized N-terminal acetyltransferase (NAT) specificities, and emerging evidence of posttranslational Nt-acetylation, necessitate the use of genetic models and quantitative proteomics. NatB, which targets Met-Glu-, Met-Asp-, and Met-Asn-starting protein N termini, is presumed to Nt-acetylate 15\% of all yeast and 18\% of all human proteins. We here report on the evolutionary traits of NatB from yeast to human and demonstrate that ectopically expressed hNatB in a yNatB-Delta yeast strain partially complements the natB-Delta phenotypes and partially restores the yNatB Nt-acetylome. Overall, combining quantitative N-terminomics with yeast studies and knockdown of hNatB in human cell lines, led to the unambiguous identification of 180 human and 110 yeast NatB substrates. Interestingly, these substrates included Met-Gln- N-termini, which are thus now classified as in vivo NatB substrates. We also demonstrate the requirement of hNatB activity for maintaining the structure and function of actomyosin fibers and for proper cellular migration. In addition, expression of tropomyosin-1 restored the altered focal adhesions and cellular migration defects observed in hNatB-depleted HeLa cells, indicative for the conserved link between NatB, tropomyosin, and actin cable function from yeast to human.},
  author       = {Van Damme, Petra and Lasa, Marta and Polevoda, Bogdan and Gazquez, Cristina and Elosegui-Artola, Alberto and Kim, Duk Soo and De Juan-Pardo, Elena and Demeyer, Kimberly and Hole, Kristine and Larrea, Esther and Timmerman, Evy and Prieto, Jesus and Arnesen, Thomas and Sherman, Fred and Gevaert, Kris and Aldabe, Rafael},
  issn         = {0027-8424},
  journal      = {PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA},
  keyword      = {ACTIN,IDENTIFICATION,TROPOMYOSIN,ACETYLATION,YEAST,CELL-MIGRATION,PROTEIN TERMINI,SACCHAROMYCES-CEREVISIAE,METHIONINE AMINOPEPTIDASE,ALPHA-ACETYLTRANSFERASE},
  language     = {eng},
  number       = {31},
  pages        = {12449--12454},
  title        = {N-terminal acetylome analyses and functional insights of the N-terminal acetyltransferase NatB},
  url          = {http://dx.doi.org/10.1073/pnas.1210303109},
  volume       = {109},
  year         = {2012},
}

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