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PER1 Modulates SGLT1 Transcription In Vitro Independent of E-box Status

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Abstract

Background and Aims

The intestine demonstrates profound circadian rhythmicity in glucose absorption in rodents, mediated entirely by rhythmicity in the transcription, translation, and function of the sodium glucose co-transporter SGLT1 (Slc5a1). Clock genes are rhythmic in the intestine and have been implicated in the regulation of rhythmicity of other intestinal genes; however, their role in the regulation of SGLT1 is unknown. We investigated the effects of one clock gene, PER1, on SGLT1 transcription in vitro.

Methods

Caco-2 cells were stably transfected with knockdown vectors for PER1 and mRNA expression of clock genes and SGLT1 determined using quantitative polymerase chain reaction (qPCR). Chinese hamster ovary (CHO) cells were transiently cotransfected with combinations of the PER1 expression vectors and the wild-type SGLT1-luciferase promoter construct or the promoter with mutated E-box sequences.

Results

Knockdown of PER1 increased native SGLT1 expression in Caco-2 enterocytes, while promoter studies confirmed that the inhibitory activity of PER1 on SGLT1 occurs via the proximal 1 kb of the SGLT1 promoter. E-box sites exerted a suppressive effect on the SGLT1 promoter; however, mutation of E-boxes had little effect on the inhibitory activity of PER1 on the SGLT1 promoter suggesting that the actions of PER1 on SGLT1 are independent of E-boxes.

Conclusions

Our findings suggest that PER1 exerts an indirect suppressive effect on SGLT1, possibly acting via other clock-controlled genes binding to non-E-box sites on the SGLT1 promoter. Understanding the regulation of rhythmicity of SGLT1 may lead to new treatments for the modulation of SGLT1 expression in conditions such as malabsorption, diabetes, and obesity.

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Abbreviations

Bmal1:

Brain muscle Arnt-like 1

Cry:

Cryptochrome

Mdr1:

Multidrug resistance 1

mRNA:

Messenger ribonucleic acid

Nhe3:

Na+/H+ exchanger

Per:

Period

PVDF:

Polyvinylidenefluoride

SGLT1:

Sodium glucose cotransporter 1

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Acknowledgments

This study is supported by NIH—5 R01 DK047326 (S.W.A.), ADA—7-05-RA-121 (D.B.R.), Harvard Clinical Nutrition Research Center—P30-DK040561 (A.T.), Nutricia Research Foundation (A.B.) and Berkeley Fellowship (A.T.S.).

Conflict of interest

All authors confirm no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Surgery, Brigham and Women’s Hospital, 75 Francis Street, Boston, MA, 02115, USA

    Anita Balakrishnan, Adam T. Stearns, Stanley W. Ashley & Ali Tavakkolizadeh

  2. Department of Surgery, Harvard Medical School, Boston, MA, 02115, USA

    Anita Balakrishnan, Adam T. Stearns, Stanley W. Ashley, David B. Rhoads & Ali Tavakkolizadeh

  3. Division of Gastroenterology, School of Clinical Sciences, University of Liverpool, Liverpool, L69 3GE, UK

    Anita Balakrishnan

  4. Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, OX1 2JD, UK

    Adam T. Stearns

  5. Pediatric Endocrine Unit, Mass General Hospital for Children, Boston, MA, 02114, USA

    David B. Rhoads

  6. Thorn 1503, Brigham and Women’s Hospital, 20 Shattuck Street, Boston, MA, 02115, USA

    Anita Balakrishnan

Authors
  1. Anita Balakrishnan
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  2. Adam T. Stearns
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  3. Stanley W. Ashley
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  5. Ali Tavakkolizadeh
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Corresponding authors

Correspondence to Anita Balakrishnan or Ali Tavakkolizadeh.

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Balakrishnan, A., Stearns, A.T., Ashley, S.W. et al. PER1 Modulates SGLT1 Transcription In Vitro Independent of E-box Status. Dig Dis Sci 57, 1525–1536 (2012). https://doi.org/10.1007/s10620-012-2166-8

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  • DOI: https://doi.org/10.1007/s10620-012-2166-8

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