tetano
Editor, Senior Moderator
FEBS J
. 2021 Jul 16.
doi: 10.1111/febs.16123. Online ahead of print.
Acetylation of the influenza A virus polymerase subunit PA in the N-terminal domain positively regulates its endonuclease activity
Dai Hatakeyama[SUP] 1 [/SUP], Masaki Shoji[SUP] 1 [/SUP], Seiryo Ogata[SUP] 2 [/SUP], Takeshi Masuda[SUP] 2 [/SUP], Masahiro Nakano[SUP] 3 [/SUP], Tsugunori Komatsu[SUP] 1 [/SUP], Ayaka Saitoh[SUP] 1 [/SUP], Kyoko Makiyama[SUP] 1 [/SUP], Hazuki Tsuneishi[SUP] 1 [/SUP], Asuka Miyatake[SUP] 1 [/SUP], Mizuki Takahira[SUP] 1 [/SUP], Erina Nishikawa[SUP] 1 [/SUP], Ayana Ohkubo[SUP] 1 [/SUP], Takeshi Noda[SUP] 3 [/SUP], Yoshihiro Kawaoka[SUP] 4 5 [/SUP], Sumio Ohtsuki[SUP] 2 [/SUP], Takashi Kuzuhara[SUP] 1 [/SUP]
Affiliations
Abstract
The post-translational acetylation of lysine residues is found in many non-histone proteins and is involved in a wide range of biological processes. Recently, we showed that the nucleoprotein of the influenza A virus is acetylated by histone acetyltransferases (HATs), a phenomenon that affects viral transcription. Here, we report that the PA subunit of influenza A virus RNA-dependent RNA polymerase is acetylated by the HATs, P300/CBP-associated factor (PCAF) and general control non-derepressible 5 (GCN5), resulting in accelerated endonuclease activity. Specifically, the full-length PA subunit expressed in cultured 293T cells was found to be strongly acetylated. Moreover, the partial recombinant protein of the PA N-terminal region containing the endonuclease domain was also acetylated by PCAF and GCN5 in vitro, which facilitated its endonuclease activity. Mass spectrometry analyses identified K19 as a candidate acetylation target in the PA N-terminal region. Notably, the substitution of the lysine residue at position 19 with glutamine, a mimic of the acetyl-lysine residue, enhanced its endonuclease activity in vitro; this point mutation also accelerated influenza A virus RNA-dependent RNA polymerase activity in the cell. Our findings suggest that PA acetylation is important for the regulation of the endonuclease and RNA polymerase activities of the influenza A virus.
Keywords: RNA polymerase; acetylation; acetyltransferase; endonuclease; influenza virus.
. 2021 Jul 16.
doi: 10.1111/febs.16123. Online ahead of print.
Acetylation of the influenza A virus polymerase subunit PA in the N-terminal domain positively regulates its endonuclease activity
Dai Hatakeyama[SUP] 1 [/SUP], Masaki Shoji[SUP] 1 [/SUP], Seiryo Ogata[SUP] 2 [/SUP], Takeshi Masuda[SUP] 2 [/SUP], Masahiro Nakano[SUP] 3 [/SUP], Tsugunori Komatsu[SUP] 1 [/SUP], Ayaka Saitoh[SUP] 1 [/SUP], Kyoko Makiyama[SUP] 1 [/SUP], Hazuki Tsuneishi[SUP] 1 [/SUP], Asuka Miyatake[SUP] 1 [/SUP], Mizuki Takahira[SUP] 1 [/SUP], Erina Nishikawa[SUP] 1 [/SUP], Ayana Ohkubo[SUP] 1 [/SUP], Takeshi Noda[SUP] 3 [/SUP], Yoshihiro Kawaoka[SUP] 4 5 [/SUP], Sumio Ohtsuki[SUP] 2 [/SUP], Takashi Kuzuhara[SUP] 1 [/SUP]
Affiliations
- PMID: 34270849
- DOI: 10.1111/febs.16123
Abstract
The post-translational acetylation of lysine residues is found in many non-histone proteins and is involved in a wide range of biological processes. Recently, we showed that the nucleoprotein of the influenza A virus is acetylated by histone acetyltransferases (HATs), a phenomenon that affects viral transcription. Here, we report that the PA subunit of influenza A virus RNA-dependent RNA polymerase is acetylated by the HATs, P300/CBP-associated factor (PCAF) and general control non-derepressible 5 (GCN5), resulting in accelerated endonuclease activity. Specifically, the full-length PA subunit expressed in cultured 293T cells was found to be strongly acetylated. Moreover, the partial recombinant protein of the PA N-terminal region containing the endonuclease domain was also acetylated by PCAF and GCN5 in vitro, which facilitated its endonuclease activity. Mass spectrometry analyses identified K19 as a candidate acetylation target in the PA N-terminal region. Notably, the substitution of the lysine residue at position 19 with glutamine, a mimic of the acetyl-lysine residue, enhanced its endonuclease activity in vitro; this point mutation also accelerated influenza A virus RNA-dependent RNA polymerase activity in the cell. Our findings suggest that PA acetylation is important for the regulation of the endonuclease and RNA polymerase activities of the influenza A virus.
Keywords: RNA polymerase; acetylation; acetyltransferase; endonuclease; influenza virus.