RT Journal
A1 Wang, Yunhao
A1 Wang, Anqi
A1 Liu, Zujun
A1 Thurman, Andrew L.
A1 Powers, Linda S.
A1 Zou, Meng
A1 Zhao, Yue
A1 Hefel, Adam
A1 Li, Yunyi
A1 Zabner, Joseph
A1 Au, Kin Fai
T1 Single-molecule long-read sequencing reveals the chromatin basis of gene expression
JF Genome Research 
JO Genome Research 
YR 2019 
FD August 01 
VO 29 
IS 8 
SP 1329 
OP 1342 
DO 10.1101/gr.251116.119 
UL http://genome.cshlp.org/content/29/8/1329.abstract 
AB Genome-wide chromatin accessibility and nucleosome occupancy profiles have been widely investigated, while the long-range dynamics remain poorly studied at the single-cell level. Here, we present a new experimental approach, methyltransferase treatment followed by single-molecule long-read sequencing (MeSMLR-seq), for long-range mapping of nucleosomes and chromatin accessibility at single DNA molecules and thus achieve comprehensive-coverage characterization of the corresponding heterogeneity. MeSMLR-seq offers direct measurements of both nucleosome-occupied and nucleosome-evicted regions on a single DNA molecule, which is challenging for many existing methods. We applied MeSMLR-seq to haploid yeast, where single DNA molecules represent single cells, and thus we could investigate the combinatorics of many (up to 356) nucleosomes at long range in single cells. We illustrated the differential organization principles of nucleosomes surrounding the transcription start site for silent and actively transcribed genes, at the single-cell level and in the long-range scale. The heterogeneous patterns of chromatin status spanning multiple genes were phased. Together with single-cell RNA-seq data, we quantitatively revealed how chromatin accessibility correlated with gene transcription positively in a highly heterogeneous scenario. Moreover, we quantified the openness of promoters and investigated the coupled chromatin changes of adjacent genes at single DNA molecules during transcription reprogramming. In addition, we revealed the coupled changes of chromatin accessibility for two neighboring glucose transporter genes in response to changes in glucose concentration.