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Title: Structural determinants and cellular environment define processed actin as the sole substrate of the N-terminal acetyltransferase NAA80

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
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  1. Univ. of Bergen (Norway); Univ. of Pennsylvania, Philadelphia, PA (United States). Perelman School of Medicine
  2. Univ. of Pennsylvania, Philadelphia, PA (United States). Perelman School of Medicine
  3. Univ. of Bergen (Norway)
  4. Univ. of Bergen (Norway); Haukeland Univ. Hospital Bergen (Norway)
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N-terminal (Nt) acetylation is a major protein modification catalyzed by N-terminal acetyltransferases (NATs). Methionine acidic N termini, including actin, are cotranslationally Nt acetylated by NatB in all eukaryotes, but animal actins containing acidic N termini, are additionally posttranslationally Nt acetylated by NAA80. Actin Nt acetylation was found to regulate cytoskeletal dynamics and motility, thus making NAA80 a potential target for cell migration regulation. In this work, we developed potent and selective bisubstrate inhibitors for NAA80 and determined the crystal structure of NAA80 in complex with such an inhibitor, revealing that NAA80 adopts a fold similar to other NAT enzymes but with a more open substrate binding region. Furthermore, in contrast to most other NATs, the substrate specificity of NAA80 is mainly derived through interactions between the enzyme and the acidic amino acids at positions 2 and 3 of the actin substrate and not residues 1 and 2. A yeast model revealed that ectopic expression of NAA80 in a strain lacking NatB activity partially restored Nt acetylation of NatB substrates, including yeast actin. Thus, NAA80 holds intrinsic capacity to posttranslationally Nt acetylate NatB-type substrates in vivo. In sum, the presence of a dominant cotranslational NatB in all eukaryotes, the specific posttranslational actin methionine removal in animals, and finally, the unique structural features of NAA80 leave only the processed actins as in vivo substrates of NAA80. Together, this study reveals the molecular and cellular basis of NAA80 Nt acetylation and provides a scaffold for development of inhibitors for the regulation of cytoskeletal properties.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Organization:
National Institutes of Health (NIH); Bergen Research Foundation (BFS); Norwegian Health Authorities of Western Norway; Meltzer Foundation; Research Council of Norway; Norwegian Cancer Society
Grant/Contract Number:
T32GM071339; R35GM118090; 230865; 249843
OSTI ID:
1499772
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 115, Issue 17; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 40 works
Citation information provided by
Web of Science

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Cited By (9)

Actin's N‐terminal acetyltransferase uncovered journal July 2018
NAT6 acetylates the N‐terminus of different forms of actin journal June 2018
Pick-ya actin – a method to purify actin isoforms with bespoke key post-translational modifications journal January 2020
Does N‐Terminal Protein Acetylation Lead to Protein Degradation? journal September 2019
Actin polymerization and cell motility are affected by NAA80-mediated posttranslational N-terminal acetylation of actin journal July 2018
NAA80 is actin’s N-terminal acetyltransferase and regulates cytoskeleton assembly and cell motility journal March 2018
A novel NAA10 p.(R83H) variant with impaired acetyltransferase activity identified in two boys with ID and microcephaly journal June 2019
NATure of actin amino-terminal acetylation journal April 2018
Molecular basis for N-terminal acetylation by human NatE and its modulation by HYPK journal February 2020

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Related Subjects

59 BASIC BIOLOGICAL SCIENCES
NAA80
acetyltransferase
N-terminal acetylation
actin
inhibitor