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Systematic identification of cis-silenced genes by trans complementation

A gene’s transcriptional output is the combined product of two inputs: diffusible factors in the cellular milieu acting in trans, and chromatin state acting in cis. Here, we describe a strategy for dissecting the relative contribution of cis versus trans mechanisms to gene regulation. Referred to as...

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Detalles Bibliográficos
Autores principales: Lee, Jae Hyun, Bugarija, Branimir, Millan, Enrique J., Walton, Noah M., Gaetz, Jedidiah, Fernandes, Croydon J., Yu, Wei-Hua, Mekel-Bobrov, Nitzan, Vallender, Tammy W., Snyder, Gregory E., Xiang, Andy Peng, Lahn, Bruce T.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2640206/
https://www.ncbi.nlm.nih.gov/pubmed/19050040
http://dx.doi.org/10.1093/hmg/ddn409
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author Lee, Jae Hyun
Bugarija, Branimir
Millan, Enrique J.
Walton, Noah M.
Gaetz, Jedidiah
Fernandes, Croydon J.
Yu, Wei-Hua
Mekel-Bobrov, Nitzan
Vallender, Tammy W.
Snyder, Gregory E.
Xiang, Andy Peng
Lahn, Bruce T.
author_facet Lee, Jae Hyun
Bugarija, Branimir
Millan, Enrique J.
Walton, Noah M.
Gaetz, Jedidiah
Fernandes, Croydon J.
Yu, Wei-Hua
Mekel-Bobrov, Nitzan
Vallender, Tammy W.
Snyder, Gregory E.
Xiang, Andy Peng
Lahn, Bruce T.
author_sort Lee, Jae Hyun
collection PubMed
description A gene’s transcriptional output is the combined product of two inputs: diffusible factors in the cellular milieu acting in trans, and chromatin state acting in cis. Here, we describe a strategy for dissecting the relative contribution of cis versus trans mechanisms to gene regulation. Referred to as trans complementation, it entails fusing two disparate cell types and searching for genes differentially expressed between the two genomes of fused cells. Any differential expression can be causally attributed to cis mechanisms because the two genomes of fused cells share a single homogenized milieu in trans. This assay uncovered a state of transcriptional competency that we termed ‘occluded’ whereby affected genes are silenced by cis-acting mechanisms in a manner that blocks them from responding to the trans-acting milieu of the cell. Importantly, occluded genes in a given cell type tend to include master triggers of alternative cell fates. Furthermore, the occluded state is maintained during cell division and is extraordinarily stable under a wide range of physiological conditions. These results support the model that the occlusion of lineage-inappropriate genes is a key mechanism of cell fate restriction. The identification of occluded genes by our assay provides a hitherto unavailable functional readout of chromatin state that is distinct from and complementary to gene expression status.
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spelling pubmed-26402062009-02-25 Systematic identification of cis-silenced genes by trans complementation Lee, Jae Hyun Bugarija, Branimir Millan, Enrique J. Walton, Noah M. Gaetz, Jedidiah Fernandes, Croydon J. Yu, Wei-Hua Mekel-Bobrov, Nitzan Vallender, Tammy W. Snyder, Gregory E. Xiang, Andy Peng Lahn, Bruce T. Hum Mol Genet Articles A gene’s transcriptional output is the combined product of two inputs: diffusible factors in the cellular milieu acting in trans, and chromatin state acting in cis. Here, we describe a strategy for dissecting the relative contribution of cis versus trans mechanisms to gene regulation. Referred to as trans complementation, it entails fusing two disparate cell types and searching for genes differentially expressed between the two genomes of fused cells. Any differential expression can be causally attributed to cis mechanisms because the two genomes of fused cells share a single homogenized milieu in trans. This assay uncovered a state of transcriptional competency that we termed ‘occluded’ whereby affected genes are silenced by cis-acting mechanisms in a manner that blocks them from responding to the trans-acting milieu of the cell. Importantly, occluded genes in a given cell type tend to include master triggers of alternative cell fates. Furthermore, the occluded state is maintained during cell division and is extraordinarily stable under a wide range of physiological conditions. These results support the model that the occlusion of lineage-inappropriate genes is a key mechanism of cell fate restriction. The identification of occluded genes by our assay provides a hitherto unavailable functional readout of chromatin state that is distinct from and complementary to gene expression status. Oxford University Press 2009-03-01 2008-12-02 /pmc/articles/PMC2640206/ /pubmed/19050040 http://dx.doi.org/10.1093/hmg/ddn409 Text en © 2008 The Author(s).
spellingShingle Articles
Lee, Jae Hyun
Bugarija, Branimir
Millan, Enrique J.
Walton, Noah M.
Gaetz, Jedidiah
Fernandes, Croydon J.
Yu, Wei-Hua
Mekel-Bobrov, Nitzan
Vallender, Tammy W.
Snyder, Gregory E.
Xiang, Andy Peng
Lahn, Bruce T.
Systematic identification of cis-silenced genes by trans complementation
title Systematic identification of cis-silenced genes by trans complementation
title_full Systematic identification of cis-silenced genes by trans complementation
title_fullStr Systematic identification of cis-silenced genes by trans complementation
title_full_unstemmed Systematic identification of cis-silenced genes by trans complementation
title_short Systematic identification of cis-silenced genes by trans complementation
title_sort systematic identification of cis-silenced genes by trans complementation
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2640206/
https://www.ncbi.nlm.nih.gov/pubmed/19050040
http://dx.doi.org/10.1093/hmg/ddn409
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