TY - JOUR
T1 - Methyl-lysine readers PHF20 and PHF20L1 define two distinct gene expression-regulating NSL complexes
AU - Van, Hieu T.
AU - Harkins, Peter R.
AU - Patel, Avni
AU - Jain, Abhinav K.
AU - Lu, Yue
AU - Bedford, Mark T.
AU - Santos, Margarida A.
N1 - Funding Information:
Acknowledgments—We thank all members of the MAS laboratory for technical support and helpful discussions. Core facilities were supported by the National Cancer Institute Cancer Center, National Institutes of Health support grant (grant no.: P30CA16672) and Cancer Prevention Research Institute of Texas (CPRIT) grants RP120348, RP170002, and RP170628. We thank the staff of the Mass Spectrometry Facility at the University of Texas Medical Branch for sample preparation, digestion, analysis by LC–MS, and informatics of the proteomic samples; the Mass Spectrometry Facility is supported in part by the CPRIT grant (grant no.: RP190682).
Funding Information:
Funding and additional information—This work was supported by a Center for Cancer Epigenetics Award (to H. T. V.), an Andrew Sabin Family Fellow Award, an American Society of Hematology Junior Faculty Scholar Award, and a CPRIT First-time Tenure-Track Faculty Award (grant no.: RR150039; to M. A. S.). M. A. S. is a CPRIT scholar in cancer research. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Publisher Copyright:
© 2022 THE AUTHORS.
PY - 2022/3/1
Y1 - 2022/3/1
N2 - The methyl-lysine readers plant homeodomain finger protein 20 (PHF20) and its homolog PHF20-like protein 1 (PHF20L1) are known components of the nonspecific lethal (NSL) complex that regulates gene expression through its histone acetyltransferase activity. In the current model, both PHF homologs coexist in the same NSL complex, although this was not formally tested; nor have the functions of PHF20 and PHF20L1 regarding NSL complex integrity and transcriptional regulation been investigated. Here, we perform an in-depth biochemical and functional characterization of PHF20 and PHF20L1 in the context of the NSL complex. Using mass spectrometry, genome-wide chromatin analysis, and proteindomain mapping, we identify the existence of two distinct NSL complexes that exclusively contain either PHF20 or PHF20L1. We show that the C-terminal domains of PHF20 and PHF20L1 are essential for complex formation with NSL, and the Tudor 2 domains are required for chromatin binding. The genome-wide chromatin landscape of PHF20-PHF20L1 shows that these proteins bind mostly to the same genomic regions, at promoters of highly expressed/housekeeping genes. Yet, deletion of PHF20 and PHF20L1 does not abrogate gene expression or impact the recruitment of the NSL complex to those target gene promoters, suggesting the existence of an alternative mechanism that compensates for the transcription of genes whose sustained expression is important for critical cellular functions. This work shifts the current paradigm and lays the foundation for studies on the differential roles of PHF20 and PHF20L1 in regulating NSL complex activity in physiological and diseases states.
AB - The methyl-lysine readers plant homeodomain finger protein 20 (PHF20) and its homolog PHF20-like protein 1 (PHF20L1) are known components of the nonspecific lethal (NSL) complex that regulates gene expression through its histone acetyltransferase activity. In the current model, both PHF homologs coexist in the same NSL complex, although this was not formally tested; nor have the functions of PHF20 and PHF20L1 regarding NSL complex integrity and transcriptional regulation been investigated. Here, we perform an in-depth biochemical and functional characterization of PHF20 and PHF20L1 in the context of the NSL complex. Using mass spectrometry, genome-wide chromatin analysis, and proteindomain mapping, we identify the existence of two distinct NSL complexes that exclusively contain either PHF20 or PHF20L1. We show that the C-terminal domains of PHF20 and PHF20L1 are essential for complex formation with NSL, and the Tudor 2 domains are required for chromatin binding. The genome-wide chromatin landscape of PHF20-PHF20L1 shows that these proteins bind mostly to the same genomic regions, at promoters of highly expressed/housekeeping genes. Yet, deletion of PHF20 and PHF20L1 does not abrogate gene expression or impact the recruitment of the NSL complex to those target gene promoters, suggesting the existence of an alternative mechanism that compensates for the transcription of genes whose sustained expression is important for critical cellular functions. This work shifts the current paradigm and lays the foundation for studies on the differential roles of PHF20 and PHF20L1 in regulating NSL complex activity in physiological and diseases states.
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U2 - 10.1016/j.jbc.2022.101588
DO - 10.1016/j.jbc.2022.101588
M3 - Article
C2 - 35033534
AN - SCOPUS:85125012272
SN - 0021-9258
VL - 298
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 3
M1 - 101588
ER -