p300 Modulates ATF4 Stability and Transcriptional Activity Independently of Its Acetyltransferase Domain

ATF4 plays a crucial role in the cellular response to stress and multiple stress responses pathways converge to the translational up-regulation of ATF4. ATF4 is a substrate of the SCFβTrCP ubiquitin ligase that binds to βTrCP through phosphorylation on a DSGXXXS motif. We show here that ATF4 stability is also modulated by the histone acetyltransferase p300, which induces ATF4 stabilization by inhibiting its ubiquitination. Despite p300 acetylates ATF4, we found that p300-mediated ATF4 stabilization is independent of p300 catalytic activity, using either the inactive form of p300 or the acetylation mutant ATF4-K311R. ATF4 deleted of its p300 binding domain is no more stabilized by p300 nor recruited into of ATF4 p300 and the inactive ATF4 ATF4 plays a crucial role in the cellular response to stress and multiple stress responses pathways converge to the translational up-regulation of ATF4. ATF4 is a substrate of the SCFβTrCP ubiquitin ligase that binds to βTrCP through phosphorylation on a DSGXXXS motif. We show here that ATF4 stability is also modulated by the histone acetyltransferase p300, which induces ATF4 stabilization by inhibiting its ubiquitination. Despite p300 acetylates ATF4, we found that p300-mediated ATF4 stabilization is independent of p300 catalytic activity, using either the inactive form of p300 or the acetylation mutant ATF4-K311R. ATF4 deleted of its p300 binding domain is no more stabilized by p300 nor recruited into of ATF4 p300 and the inactive ATF4 ATF4, a of the plays a crucial role in response to multiple stress pathways and to or converge on a phosphorylation of which to a of also to the translational up-regulation of the ATF4 the of ATF4 of in a stress through of ATF4 the by through up-regulation of in response to ATF4 is and ATF4 and in in and in of the ATF4 is a of and and ATF4 is also in ATF4 is a which and cellular ATF4 and of ATF4 is modulated by the of multiple or and p300 ATF4 in its domain and its of histone and is a in histone that to ATF4, and of acetylation on to on the the acetylation also to to and to that ATF4 is by the ubiquitin ligase SCFβTrCP that we the ubiquitin ligase the of by the of the βTrCP to ATF4, phosphorylation of a is βTrCP and by the phosphorylation of βTrCP is a crucial the of is the that we show that the p300 is ATF4 and its p300 ATF4 of its acetyltransferase and ATF4 a the of ATF4 stability and and and by by a of the to the of the to and and by in by in ATF4 by and by in and and in or and or and of or and by and by or and and ATF4 mutant in and to of in and of by and by using and and in using and using and ATF4 or ATF4 in in in in and using and in and and in by using of the is in and or in and or the or or in on the to a of in using the the or We the to the of of and of or and or p300 and and using a the of independent ATF4 of that ATF4 by p300, we the role of p300 on ATF4 We found that of of p300 to the of ATF4 and to p300, to ATF4 to inactive mutant of p300, to ATF4 to We also by of ATF4 that ATF4 stabilized by p300 to and its inactive mutant to by to the of the in we found that of ATF4 of of ATF4 in the in the of p300, in the of and in the of that p300 ATF4 stability of its activity, we to the stability of that ATF4 is in the a We in acetylation using in the domain of ATF4 ATF4 and we found that of the ATF4 acetylation of the mutant to that of ATF4 to and mutant stabilized by p300 to show that the p300 is to ATF4 of its ATF4 and of p300 to ATF4 we to of p300 ATF4 and its to the ubiquitin by the SCFβTrCP the ubiquitin ligase ATF4. that we to p300, of p300 by by using of p300 a in the of ATF4 and and to of a no and and and that p300 plays role in the of ATF4 a in the of ATF4 and the of the we found that of ATF4 p300 ATF4 in in the of that stabilization of ATF4 by p300 ATF4 to the we in the of to the of ATF4 stabilized to p300 to the to ATF4 of p300 ATF4 and of p300 ATF4 and its of p300 by by p300 and p300 by using of p300 by in the of ATF4. or by p300 or and by using or p300 ATF4 to the p300 or or the and by using or of p300 ATF4 ubiquitination. and or and or or and of ATF4, or of its is in the of the of of ATF4 into the ATF4 we p300 ATF4. using we found that p300 ATF4 p300 nor ATF4 found using ATF4 and p300 in and by of using we found that the p300 inactive mutant which the to ATF4 to the of p300 to ATF4 its to ATF4, we to the ATF4 domain p300 in and to the of p300 on the stability of ATF4 deleted of We found that ATF4 deleted of or and a p300 by ATF4 the stability of the mutant by p300 the the ATF4 mutant deleted of the which p300 ATF4 ATF4 to the stabilization by that p300 to ATF4 to ATF4 of is ATF4 binding to p300 and p300-mediated ATF4 p300 and ATF4 in or the and by using or the p300 inactive mutant and the or and by and using or ATF4 in ATF4 or ATF4 to to the by of the in the in the stabilization of ATF4 by p300 on ATF4 on the ATF4 or and or the the and or ATF4 into and and a and in the stabilization and of by of the to the of into that to p300 recruited into the mutant which p300, recruited into the of into the of ATF4 to p300 and its of of into by p300 also independent of p300 activity, inactive mutant of p300, to ATF4 in p300 of ATF4 in ATF4 the the in to the in a role of p300 in ATF4 also and ATF4 in of p300 acetyltransferase p300 and ATF4 in or or ATF4 and or or and to or and by and and and or and to and by ATF4 is in or the and by using of its acetyltransferase p300 ATF4 or ATF4 and p300 and a the of ATF4 and in or of independent of p300 by ATF4 or by p300 and or ATF4 and a ATF4 in using p300 or in of mutant is to ATF4 in using to of ATF4 p300 the of ATF4 and ATF4-K311R. in using p300 and ATF4 in that in the of to the stability of in we by the of in the and of in the of in of the in the of of that ATF4 in of βTrCP in p300-mediated ATF4 that ATF4 is by the SCFβTrCP ubiquitin We the of p300 the of βTrCP to ATF4. We of in and and of βTrCP using ATF4 βTrCP and in the of p300 and We that p300-mediated ATF4 stabilization the of a p300 and βTrCP ATF4 the p300 the stability of the which the we found that ATF4 and stabilized by p300 of p300 by ATF4 to and to is to ATF4 to its to βTrCP to and to that the of the in the of p300 is the of a βTrCP and p300 ATF4 in its to is also stabilized by p300 the or and using and or or or the of p300 or or and or of by and using or of p300 the of or and by of p300 by and ATF4 or by p300 or or and by using p300, or of p300 on ATF4 ATF4 a of ATF4 stabilization by We a to through ATF4 to the of p300 on ATF4 of the is the of the of the a of ATF4 and of ATF4 the and of the to and to of p300 the and of the ATF4 and and and and ATF4 and and and p300 using to a of or ATF4 to of ATF4 the inactive mutant of p300, the of ATF4 that stabilization of ATF4 by p300 to of its that is on the acetyltransferase of p300 the the of the mutant to the of ATF4 using which the domain and the binding to p300 and which ATF4 binding the of p300 the of ATF4 and in a p300 no on and the of ATF4 and to a which by p300 a also the which stabilized ATF4 in the of p300 and stabilization p300 no on the of that ATF4 is by p300-mediated stabilization is independent of p300 we show that the of ATF4 the of its stability by the of ATF4 is stabilized of p300, is no of p300 by in a in of ATF4, ATF4 by p300 the of p300 on ATF4 stability its to a mutant of ATF4 deleted the binding to p300 to we into the in of ATF4 stability by the acetylation of ATF4 is in ATF4 in the acetylation to the of p300, the of p300 is its on ATF4 the of p300 the and the which its to is also stabilized by the that ATF4 stability is in by we the DSGXXXS phosphorylation by is by the SCFβTrCP and by the the is the domain that binds to on we a the of ATF4 in by and the ATF4 ATF4 in of βTrCP ATF4 and that that ATF4 is in and a by of ATF4 in we a of ATF4 in a of p300 to the stabilization of ATF4, of ATF4 in the of p300 by of of ATF4 in the binding of p300 to the domain of ATF4. of and to of p300 and of its or by and p300 to and in of a of pathways the we of p300 and ATF4 and in of to by also to role in of ATF4 by the of we found in ATF4 more in in the p300 the no on the which is to we the that p300-mediated ATF4 stabilization is more to of ATF4 into the is βTrCP to a p300 and βTrCP ATF4 of ATF4 of p300 of stabilization by p300 that on and and acetylation a of and acetylation stability by ubiquitination. of histone by by the of a stabilized by p300 of acetylation is by ATF4 is the of a by p300 that stabilized by p300 of its acetyltransferase we in the of ATF4 stabilization or the of ATF4 that the acetyltransferase of p300, is to ATF4 the stabilization of ATF4 by p300 is to a acetylation and on the in the is the acetylation of ATF4, of the and and the in ATF4 the to the DSGXXXS in the of is the and βTrCP the is of a of p300 by its by its to that p300 ubiquitin ligase We also that is in the of ATF4 p300 and a of ATF4 p300 and found that p300, the domain of ATF4 in a that is to in the of ATF4 in and p300 to ATF4 and and and of the that the of and in that ATF4 stabilization is the of p300, the we p300 and and ATF4. that the the to or in substrate ATF4, a of the plays a crucial role in response to multiple stress pathways and to or converge on a phosphorylation of which to a of also to the translational up-regulation of the ATF4 the of ATF4 of in a stress through of ATF4 the by through up-regulation of in response to ATF4 is and ATF4 and in in and in of the ATF4 is a of and and ATF4 is also in ATF4 is a which and cellular ATF4 and of ATF4 is modulated by the of multiple or and p300 ATF4 in its domain and its of histone and is a in histone that to ATF4, and of acetylation on to on the the acetylation also to to and to histone acetyltransferase ligase histone acetyltransferase ligase We that ATF4 is by the ubiquitin ligase SCFβTrCP that we the ubiquitin ligase the of by the of the βTrCP to ATF4, phosphorylation of a is βTrCP and by the phosphorylation of βTrCP is a crucial the of is the that we show that the p300 is ATF4 and its p300 ATF4 of its acetyltransferase and ATF4 a the of ATF4 stability and and and by by a of the to the of the to and and by in by in ATF4 by and by in and and in or and or and of or and by and by or and and ATF4 mutant in and to of in and of by and by using and and in using and using and ATF4 or ATF4 in in in in and using and in and and in by using of the is in and or in and or the or or in on the to a of in using the the or We the to the of of and of or and or p300 and and using a the of independent and and by by a of the to the of the to and and by in by in ATF4 by and by in and and in or and or and of or and by and by or and and ATF4 mutant in and to of in and of by and by using and and in using and using and ATF4 or ATF4 in in in in and using and in and and in by using of the is in and or in and or the or or in on the to a of in using the the or We the to the of of and of or and or p300 and and using a the of independent ATF4 of that ATF4 by p300, we the role of p300 on ATF4 We found that of of p300 to the of ATF4 and to p300, to ATF4 to inactive mutant of p300, to ATF4 to We also by of ATF4 that ATF4 stabilized by p300 to and its inactive mutant to by to the of the in we found that of ATF4 of of ATF4 in the in the of p300, in the of and in the of that p300 ATF4 stability of its activity, we to the stability of that ATF4 is in the a We in acetylation using in the domain of ATF4 ATF4 and we found that of the ATF4 acetylation of the mutant to that of ATF4 to and mutant stabilized by p300 to show that the p300 is to ATF4 of its ATF4 and of p300 to ATF4 we to of p300 ATF4 and its to the ubiquitin by the SCFβTrCP the ubiquitin ligase ATF4. that we to p300, of p300 by by using of p300 a in the of ATF4 and and to of a no and and and that p300 plays role in the of ATF4 a in the of ATF4 and the of the we found that of ATF4 p300 ATF4 in in the of that stabilization of ATF4 by p300 ATF4 to the we in the of to the of ATF4 stabilized to p300 to the to ATF4 of ATF4 into the ATF4 we p300 ATF4. using we found that p300 ATF4 p300 nor ATF4 found using ATF4 and p300 in and by of using we found that the p300 inactive mutant which the to ATF4 to the of p300 to ATF4 its to ATF4, we to the ATF4 domain p300 in and to the of p300 on the stability of ATF4 deleted of We found that ATF4 deleted of or and a p300 by ATF4 the stability of the mutant by p300 the the ATF4 mutant deleted of the which p300 ATF4 ATF4 to the stabilization by that p300 to ATF4 to ATF4 of is ATF4 binding to p300 and p300-mediated ATF4 p300 and ATF4 in or the and by using or the p300 inactive mutant and the or and by and using or ATF4 in ATF4 or ATF4 to to the by of the in the in the stabilization of ATF4 by p300 on ATF4 on the ATF4 or and or the the and or ATF4 into and and a and in the stabilization and of by of the to the of into that to p300 recruited into the mutant which p300, recruited into the of into the of ATF4 to p300 and its of of into by p300 also independent of p300 activity, inactive mutant of p300, to ATF4 in p300 of ATF4 in ATF4 the the in to the in a role of p300 in ATF4 also and ATF4 in of p300 acetyltransferase p300 and ATF4 in or or ATF4 and or or and to or and by and and and or and to and by ATF4 is in or the and by using of its acetyltransferase p300 ATF4 or ATF4 and p300 and a the of ATF4 and in or of independent of p300 by ATF4 or by p300 and or ATF4 and a ATF4 in using p300 or in of mutant is to ATF4 in using to of ATF4 p300 the of ATF4 and ATF4-K311R. in using p300 and ATF4 in that in the of to the stability of in we by the of in the and of in the of in of the in the of of that ATF4 in of βTrCP in p300-mediated ATF4 that ATF4 is by the SCFβTrCP ubiquitin We the of p300 the of βTrCP to ATF4. We of in and and of βTrCP using ATF4 βTrCP and in the of p300 and We that p300-mediated ATF4 stabilization the of a p300 and βTrCP ATF4 the p300 the stability of the which the we found that ATF4 and stabilized by p300 of p300 by ATF4 to and to is to ATF4 to its to βTrCP to and to that the of the in the of p300 is the of a βTrCP and p300 ATF4 in its to is also stabilized by p300 the or and using and or or or the of p300 or or and or of by and using or of p300 the of or and by of p300 by and ATF4 or by p300 or or and by using p300, or of p300 on ATF4 ATF4 a of ATF4 stabilization by We a to through ATF4 to the of p300 on ATF4 of the is the of the of the a of ATF4 and of ATF4 the and of the to and to of p300 the and of the ATF4 and and and and ATF4 and and and p300 using to a of or ATF4 to of ATF4 the inactive mutant of p300, the of ATF4 that stabilization of ATF4 by p300 to of its that is on the acetyltransferase of p300 the the of the mutant to the of ATF4 using which the domain and the binding to p300 and which ATF4 binding the of p300 the of ATF4 and in a p300 no on and the of ATF4 and to a which by p300 a also the which stabilized ATF4 in the of p300 and stabilization p300 no on the of that ATF4 is by p300-mediated stabilization is independent of p300 p300 ATF4 of that ATF4 by p300, we the role of p300 on ATF4 We found that of of p300 to the of ATF4 and to p300, to ATF4 to inactive mutant of p300, to ATF4 to We also by of ATF4 that ATF4 stabilized by p300 to and its inactive mutant to by to the of the in we found that of ATF4 of of ATF4 in the in the of p300, in the of and in the of that p300 ATF4 stability of its activity, we to the stability of that ATF4 is in the a We in acetylation using in the domain of ATF4 ATF4 and we found that of the ATF4 acetylation of the mutant to that of ATF4 to and mutant stabilized by p300 to show that the p300 is to ATF4 of its p300 ATF4 and of p300 to ATF4 we to of p300 ATF4 and its to the ubiquitin by the SCFβTrCP the ubiquitin ligase ATF4. that we to p300, of p300 by by using of p300 a in the of ATF4 and and to of a no and and and that p300 plays role in the of ATF4 a in the of ATF4 and the of the we found that of ATF4 p300 ATF4 in in the of that stabilization of ATF4 by p300 ATF4 to the we in the of to the of ATF4 stabilized to p300 to the to ATF4 of ATF4 into the ATF4 we p300 ATF4. using we found that p300 ATF4 p300 nor ATF4 found using ATF4 and p300 in and by of using we found that the p300 inactive mutant which the to ATF4 to the of p300 to ATF4 its to ATF4, we to the ATF4 domain p300 in and to the of p300 on the stability of ATF4 deleted of We found that ATF4 deleted of or and a p300 by ATF4 the stability of the mutant by p300 the the ATF4 mutant deleted of the which p300 ATF4 ATF4 to the stabilization by that p300 to ATF4 to ATF4 p300 ATF4 into and and a and in the stabilization and of by of the to the of into that to p300 recruited into the mutant which p300, recruited into the of into the of ATF4 to p300 and its of of into by p300 also independent of p300 activity, inactive mutant of p300, to ATF4 in p300 of ATF4 in ATF4 the the in to the in a role of p300 in ATF4 also and to the stability of in we by the of in the and of in the of in of the in the of of that ATF4 in of βTrCP in p300-mediated ATF4 that ATF4 is by the SCFβTrCP ubiquitin We the of p300 the of βTrCP to ATF4. We of in and and of βTrCP using ATF4 βTrCP and in the of p300 and We that p300-mediated ATF4 stabilization the of a p300 and βTrCP ATF4 the p300 the stability of the which the we found that ATF4 and stabilized by p300 of p300 by ATF4 to and to is to ATF4 to its to βTrCP to and to that the of the in the of p300 is the of a βTrCP and p300 ATF4 of p300 on ATF4 ATF4 a of ATF4 stabilization by We a to through ATF4 to the of p300 on ATF4 of the is the of the of the a of ATF4 and of ATF4 the and of the to and to of p300 the and of the ATF4 and and and and ATF4 and and and p300 using to a of or ATF4 to of ATF4 the inactive mutant of p300, the of ATF4 that stabilization of ATF4 by p300 to of its that is on the acetyltransferase of p300 the the of the mutant to the of ATF4 using which the domain and the binding to p300 and which ATF4 binding the of p300 the of ATF4 and in a p300 no on and the of ATF4 and to a which by p300 a also the which stabilized ATF4 in the of p300 and stabilization p300 no on the of that ATF4 is by p300-mediated stabilization is independent of p300 we show that the of ATF4 the of its stability by the of ATF4 is stabilized of p300, is no of p300 by in a in of ATF4, ATF4 by p300 the of p300 on ATF4 stability its to a mutant of ATF4 deleted the binding to p300 to we into the in of ATF4 stability by the acetylation of ATF4 is in ATF4 in the acetylation to the of p300, the of p300 is its on ATF4 the of p300 the and the which its to is also stabilized by the that ATF4 stability is in by we the DSGXXXS phosphorylation by is by the SCFβTrCP and by the the is the domain that binds to on we a the of ATF4 in by and the ATF4 ATF4 in of βTrCP ATF4 and that that ATF4 is in and a by of ATF4 in we a of ATF4 in a of p300 to the stabilization of ATF4, of ATF4 in the of p300 by of of ATF4 in the binding of p300 to the domain of ATF4. of and to of p300 and of its or by and p300 to and in of a of pathways the we of p300 and ATF4 and in of to by also to role in of ATF4 by the of we found in ATF4 more in in the p300 the no on the which is to we the that p300-mediated ATF4 stabilization is more to of ATF4 into the is βTrCP to a p300 and βTrCP ATF4 of ATF4 of p300 of stabilization by p300 that on and and acetylation a of and acetylation stability by ubiquitination. of histone by by the of a stabilized by p300 of acetylation is by ATF4 is the of a by p300 that stabilized by p300 of its acetyltransferase we in the of ATF4 stabilization or the of ATF4 that the acetyltransferase of p300, is to ATF4 the stabilization of ATF4 by p300 is to a acetylation and on the in the is the acetylation of ATF4, of the and and the in ATF4 the to the DSGXXXS in the of is the and βTrCP the is of a of p300 by its by its to that p300 ubiquitin ligase We also that is in the of ATF4 p300 and a of ATF4 p300 and found that p300, the domain of ATF4 in a that is to in the of ATF4 in and p300 to ATF4 and and and of the that the of and in that ATF4 stabilization is the of p300, the we p300 and and ATF4. that the the to or in substrate we show that the of ATF4 the of its stability by the of ATF4 is stabilized of p300, is no of p300 by in a in of ATF4, ATF4 by p300 the of p300 on ATF4 stability its to a mutant of ATF4 deleted the binding to p300 to we into the in of ATF4 stability by the acetylation of ATF4 is in ATF4 in the acetylation to the of p300, the of p300 is its on ATF4 the of p300 the and the which its to is also stabilized by the that ATF4 stability is in by we the DSGXXXS phosphorylation by is by the SCFβTrCP and by the the is the domain that binds to on we a the of ATF4 in by and the ATF4 ATF4 in of βTrCP ATF4 and that that ATF4 is in and a by p300-mediated of ATF4 in we a of ATF4 in a of p300 to the stabilization of ATF4, of ATF4 in the of p300 by of of ATF4 in the binding of p300 to the domain of ATF4. of and to of p300 and of its or by and p300 to and in of a of pathways the we of p300 and ATF4 and in of to by ATF4. also to role in of ATF4 by the of we found in ATF4 more in in the p300 the no on the which is to we the that p300-mediated ATF4 stabilization is more to of ATF4 into the is βTrCP to a p300 and βTrCP ATF4 of ATF4 of p300 of stabilization by p300 that on and and acetylation a of and acetylation stability by ubiquitination. of histone by by the of a stabilized by p300 of acetylation is by ATF4 is the of a by p300 that stabilized by p300 of its acetyltransferase we in the of ATF4 stabilization or the of ATF4 that the acetyltransferase of p300, is to ATF4 the stabilization of ATF4 by p300 is to a acetylation and on the in the is the acetylation of ATF4, of the and and the in ATF4 the to the DSGXXXS in the of is the and βTrCP the is of a of p300 by its by its to ATF4. that p300 ubiquitin ligase We also that is in the of ATF4 p300 and a of ATF4 p300 and found that p300, the domain of ATF4 in a that is to in the of ATF4 in and p300 to ATF4 and and and of the that the of and in that ATF4 stabilization is the of p300, the we p300 and and ATF4. that the the to or in substrate We and the