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Chromosome Research

, Volume 26, Issue 4, pp 255–276 | Cite as

Nuclear localization signal region in nuclear receptor PXR governs the receptor association with mitotic chromatin

  • Manjul Rana
  • Amit K. Dash
  • Kalaiarasan Ponnusamy
  • Rakesh K. TyagiEmail author
Original Article

Abstract

In recent years, some transcription factors have been observed to remain associated with mitotic chromatin. Based on these observations, it is suggested that these chromatin-bound transcription factors may serve as ‘epigenetic marks’ for transmission of pattern of gene expression from progenitor to progeny cells. In this context, our laboratory has reported that nuclear receptor PXR, a master regulator of xenobiotic metabolism, remains constitutively associated with mitotic chromatin. However, the region responsible for this interaction with chromatin remained unknown. In this study, we have shown, for the first time, that mitotic chromatin association of this factor is mediated by the combined action of two zinc fingers present in the DNA-binding domain of PXR. Overall, the nuclear localization signal (NLS) region appears to play a major role in this interaction with mitotic chromatin. Also, we have identified a sub-region of 11 amino acid residues within NLS region of PXR (R66-76R) essential for receptor interaction with the mitotic chromatin. Interestingly, this minimal region is sequence-specific and independent of its basic charge. We have termed this minimal sub-region as ‘mitotic chromatin binding-determining region’ (MCBR). It is suggested that this receptor region is essential for activation of its target genes. Additionally, we have shown that PXR remains associated with the everted repeat (ER6) region of its major target gene, CYP3A4 promoter during mitosis implying its suggested role in ‘gene bookmarking’.

Keywords

Nuclear receptors Pregnane and xenobiotic receptor Mitosis Zinc fingers Nuclear localization signal Gene bookmarking 

Abbreviations

AR

Androgen receptor

BIOPIT

Bimolecular imprints offered to progeny for inheritance of traits

Cdks

Cyclin-dependent kinases

ChIP

Chromatin immunoprecipitation

CTCF

CCCTC-binding factor

DHT

Dihydrotestosterone

ER6

Everted repeat 6

ERα

Estrogen receptor α

Esrrb

Estrogen-related receptor beta

FACS

Fluorescence-activated cell sorting

FLIM

Fluorescence-lifetime imaging microscopy

FOX1

Forkhead box 1

FRET

Fluorescence resonance energy transfer

HMGA1a

High-mobility group proteins A1

HMGB1

High-mobility group box 1

HMGB2

High-mobility group protein B2

hsp70

70 kilodalton heat shock proteins

MCBR

Mitotic chromatin binding-determining region

NLS

Nuclear localization signal

PARP-1

Poly (ADP-ribose) polymerase-1

PcG proteins

Polycomb-group proteins

PXR

Pregnane and xenobiotic receptor

RBPJ

Recombining binding protein suppressor of hairless

Runx3

Runt-related transcription factor 3

TBP

TATA-box-binding protein

TFIID

Transcription factor II D

TFIIH

Transcription factor II Human

XREM

Xenobiotic response element

Notes

Acknowledgements

We acknowledge Dr. Amulya K. Panda, Director, and Dr. Vikash Kumar, Scientist at the National Institute of Immunology, New Delhi, for help with the cell cycle analysis and sorting of mitotic cells.

Authors’ contributions

RKT conceived, designed, and supervised the project. MR and AKD designed and performed most of the experiments. RKT, MR, and AKD analyzed the data and wrote the paper. KP did the in silico experiments.

Funding

The research work presented in this paper was financially supported by a research grant to RKT from the UPE-II (University with Potential for Excellence phase II)- project ID 25 and UGC (University Grants Commission)- major project - F.No. 41-1294/2012(SR), Central financial support to our Centre by UGC-SAP (University Grants Commission-Special Assistance Programme) - F.No. 3-17/2015/DRS II (SAP-II), ICMR-CAR (Indian Council of Medical Research-Centre for Advanced Research) - F.No. 63/9/2010-BMS, DST-PURSE (Department of Science & Technology-Promotion of University Research and Scientific Excellence) - PAC-JNU-DST-PURSE-462 (Phase-II) is gratefully acknowledged. MR, AKD, and KP acknowledge UGC-BSR (University Grants Commission - Basic Scientific Research), DST-INSPIRE (Department of Science & Technology- Innovation in Science Pursuit for Inspired Research) and DST-SERB (Department of Science & Technology-Science and Engineering Research Board) respectively for the grant of doctoral research fellowships (MR and AKD) and post-doctoral research fellowship (KP).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

10577_2018_9583_MOESM1_ESM.doc (10.9 mb)
ESM 1 (DOC 11140 kb)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Manjul Rana
    • 1
  • Amit K. Dash
    • 1
  • Kalaiarasan Ponnusamy
    • 2
  • Rakesh K. Tyagi
    • 1
    Email author
  1. 1.Special Centre for Molecular MedicineJawaharlal Nehru UniversityNew DelhiIndia
  2. 2.Synthetic Biology Lab, School of BiotechnologyJawaharlal Nehru UniversityNew DelhiIndia

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