Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells

Prachi N. Ghule, Zbigniew Dominski, Xiao Cui Yang, William F. Marzluff, Klaus A. Becker, J. Wade Harper, Jane B. Lian, Janet L. Stein, Andre J van Wijnen, Gary S. Stein

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Human embryonic stem (hES) cells have an abbreviated G1 phase of the cell cycle. How cells expedite G1 events that are required for the initiation of S phase has not been resolved. One key regulatory pathway that controls G1/S-phase transition is the cyclin E/CDK2-dependent activation of the coactivator protein nuclear protein, ataxia-telangiectasia locus/histone nuclear factor-P (p220NPAT/HiNF-P) complex that induces histone gene transcription. In this study, we use the subnuclear organization of factors controlling histone gene expression to define mechanistic differences in the G1 phase of hES and somatic cells using in situ immunofluorescence microscopy and fluorescence in situ hybridization (FISH). We show that histone gene expression is supported by the staged assembly and modification of a unique subnuclear structure that coordinates initiation and processing of transcripts originating from histone gene loci. Our results demonstrate that regulatory complexes that mediate transcriptional initiation (e.g., p220NPAT) and 3′-end processing (e.g., Lsm10, Lsm11, and SLBP) of histone gene transcripts colocalize at histone gene loci in dedicated subnuclear foci (histone locus bodies) that are distinct from Cajal bodies. Although appearance of CDK2-phosphorylated p220NPAT in these domains occurs at the time of S-phase entry, histone locus bodies are formed ≈1 to 2 h before S phase in embryonic cells but 6 h before S phase in somatic cells. These temporal differences in the formation of histone locus bodies suggest that the G1 phase of the cell cycle in hES cells is abbreviated in part by contraction of late G1.

Original languageEnglish (US)
Pages (from-to)16964-16969
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume105
Issue number44
DOIs
StatePublished - Nov 4 2008
Externally publishedYes

Fingerprint

Histones
Cell Cycle
Gene Expression
S Phase
G1 Phase
Genes
Human Embryonic Stem Cells
Cyclin E
Ataxia Telangiectasia
Phase Transition
Nuclear Proteins
Fluorescence In Situ Hybridization
Fluorescence Microscopy

Keywords

  • Cajal body
  • Coilin
  • G1/S transition
  • HiNF-P
  • p220

ASJC Scopus subject areas

  • General

Cite this

Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells. / Ghule, Prachi N.; Dominski, Zbigniew; Yang, Xiao Cui; Marzluff, William F.; Becker, Klaus A.; Harper, J. Wade; Lian, Jane B.; Stein, Janet L.; van Wijnen, Andre J; Stein, Gary S.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, No. 44, 04.11.2008, p. 16964-16969.

Research output: Contribution to journalArticle

Ghule, PN, Dominski, Z, Yang, XC, Marzluff, WF, Becker, KA, Harper, JW, Lian, JB, Stein, JL, van Wijnen, AJ & Stein, GS 2008, 'Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells', Proceedings of the National Academy of Sciences of the United States of America, vol. 105, no. 44, pp. 16964-16969. https://doi.org/10.1073/pnas.0809273105
Ghule, Prachi N. ; Dominski, Zbigniew ; Yang, Xiao Cui ; Marzluff, William F. ; Becker, Klaus A. ; Harper, J. Wade ; Lian, Jane B. ; Stein, Janet L. ; van Wijnen, Andre J ; Stein, Gary S. / Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells. In: Proceedings of the National Academy of Sciences of the United States of America. 2008 ; Vol. 105, No. 44. pp. 16964-16969.
@article{d7ef2b9c565c452b8e614083f3ce1cdf,
title = "Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells",
abstract = "Human embryonic stem (hES) cells have an abbreviated G1 phase of the cell cycle. How cells expedite G1 events that are required for the initiation of S phase has not been resolved. One key regulatory pathway that controls G1/S-phase transition is the cyclin E/CDK2-dependent activation of the coactivator protein nuclear protein, ataxia-telangiectasia locus/histone nuclear factor-P (p220NPAT/HiNF-P) complex that induces histone gene transcription. In this study, we use the subnuclear organization of factors controlling histone gene expression to define mechanistic differences in the G1 phase of hES and somatic cells using in situ immunofluorescence microscopy and fluorescence in situ hybridization (FISH). We show that histone gene expression is supported by the staged assembly and modification of a unique subnuclear structure that coordinates initiation and processing of transcripts originating from histone gene loci. Our results demonstrate that regulatory complexes that mediate transcriptional initiation (e.g., p220NPAT) and 3′-end processing (e.g., Lsm10, Lsm11, and SLBP) of histone gene transcripts colocalize at histone gene loci in dedicated subnuclear foci (histone locus bodies) that are distinct from Cajal bodies. Although appearance of CDK2-phosphorylated p220NPAT in these domains occurs at the time of S-phase entry, histone locus bodies are formed ≈1 to 2 h before S phase in embryonic cells but 6 h before S phase in somatic cells. These temporal differences in the formation of histone locus bodies suggest that the G1 phase of the cell cycle in hES cells is abbreviated in part by contraction of late G1.",
keywords = "Cajal body, Coilin, G1/S transition, HiNF-P, p220",
author = "Ghule, {Prachi N.} and Zbigniew Dominski and Yang, {Xiao Cui} and Marzluff, {William F.} and Becker, {Klaus A.} and Harper, {J. Wade} and Lian, {Jane B.} and Stein, {Janet L.} and {van Wijnen}, {Andre J} and Stein, {Gary S.}",
year = "2008",
month = "11",
day = "4",
doi = "10.1073/pnas.0809273105",
language = "English (US)",
volume = "105",
pages = "16964--16969",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "44",

}

TY - JOUR

T1 - Staged assembly of histone gene expression machinery at subnuclear foci in the abbreviated cell cycle of human embryonic stem cells

AU - Ghule, Prachi N.

AU - Dominski, Zbigniew

AU - Yang, Xiao Cui

AU - Marzluff, William F.

AU - Becker, Klaus A.

AU - Harper, J. Wade

AU - Lian, Jane B.

AU - Stein, Janet L.

AU - van Wijnen, Andre J

AU - Stein, Gary S.

PY - 2008/11/4

Y1 - 2008/11/4

N2 - Human embryonic stem (hES) cells have an abbreviated G1 phase of the cell cycle. How cells expedite G1 events that are required for the initiation of S phase has not been resolved. One key regulatory pathway that controls G1/S-phase transition is the cyclin E/CDK2-dependent activation of the coactivator protein nuclear protein, ataxia-telangiectasia locus/histone nuclear factor-P (p220NPAT/HiNF-P) complex that induces histone gene transcription. In this study, we use the subnuclear organization of factors controlling histone gene expression to define mechanistic differences in the G1 phase of hES and somatic cells using in situ immunofluorescence microscopy and fluorescence in situ hybridization (FISH). We show that histone gene expression is supported by the staged assembly and modification of a unique subnuclear structure that coordinates initiation and processing of transcripts originating from histone gene loci. Our results demonstrate that regulatory complexes that mediate transcriptional initiation (e.g., p220NPAT) and 3′-end processing (e.g., Lsm10, Lsm11, and SLBP) of histone gene transcripts colocalize at histone gene loci in dedicated subnuclear foci (histone locus bodies) that are distinct from Cajal bodies. Although appearance of CDK2-phosphorylated p220NPAT in these domains occurs at the time of S-phase entry, histone locus bodies are formed ≈1 to 2 h before S phase in embryonic cells but 6 h before S phase in somatic cells. These temporal differences in the formation of histone locus bodies suggest that the G1 phase of the cell cycle in hES cells is abbreviated in part by contraction of late G1.

AB - Human embryonic stem (hES) cells have an abbreviated G1 phase of the cell cycle. How cells expedite G1 events that are required for the initiation of S phase has not been resolved. One key regulatory pathway that controls G1/S-phase transition is the cyclin E/CDK2-dependent activation of the coactivator protein nuclear protein, ataxia-telangiectasia locus/histone nuclear factor-P (p220NPAT/HiNF-P) complex that induces histone gene transcription. In this study, we use the subnuclear organization of factors controlling histone gene expression to define mechanistic differences in the G1 phase of hES and somatic cells using in situ immunofluorescence microscopy and fluorescence in situ hybridization (FISH). We show that histone gene expression is supported by the staged assembly and modification of a unique subnuclear structure that coordinates initiation and processing of transcripts originating from histone gene loci. Our results demonstrate that regulatory complexes that mediate transcriptional initiation (e.g., p220NPAT) and 3′-end processing (e.g., Lsm10, Lsm11, and SLBP) of histone gene transcripts colocalize at histone gene loci in dedicated subnuclear foci (histone locus bodies) that are distinct from Cajal bodies. Although appearance of CDK2-phosphorylated p220NPAT in these domains occurs at the time of S-phase entry, histone locus bodies are formed ≈1 to 2 h before S phase in embryonic cells but 6 h before S phase in somatic cells. These temporal differences in the formation of histone locus bodies suggest that the G1 phase of the cell cycle in hES cells is abbreviated in part by contraction of late G1.

KW - Cajal body

KW - Coilin

KW - G1/S transition

KW - HiNF-P

KW - p220

UR - http://www.scopus.com/inward/record.url?scp=55949109449&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=55949109449&partnerID=8YFLogxK

U2 - 10.1073/pnas.0809273105

DO - 10.1073/pnas.0809273105

M3 - Article

C2 - 18957539

AN - SCOPUS:55949109449

VL - 105

SP - 16964

EP - 16969

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 44

ER -