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TP53 status regulates ACSL5-induced expression of mitochondrial mortalin in enterocytes and colorectal adenocarcinomas

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Abstract

Acyl-CoA synthetase 5 (ACSL5), a mitochondrially localized enzyme, catalyzes the synthesis of long-chain fatty acid thioesters and is physiologically involved in pro-apoptotic sensing of enterocytes. The aim of the present study is to identify an ACSL5-dependent regulation of mitochondrially expressed proteins and the characterization of related pathways in normal and diseased human intestinal mucosa. Proteomics of isolated mitochondria from ACSL5 transfectants and CaCo2 controls were performed. ACSL5-dependent protein synthesis was verified with quantitative reverse transcription plus the polymerase chain reaction, Western blotting, short-interfering-RNA-mediated gene silencing and additional cell culture experiments. Lipid changes were analyzed with tandem mass spectrometry. ACSL5-related pathways were characterized in normal mucosa and sporadic adenocarcinomas of the human intestine. In CaCo2 cells transfected with ACSL5, mortalin (HSPA9) was about two-fold increased in mitochondria, whereas cytoplasmic mortalin levels were unchanged. Disturbance of acyl-CoA/sphingolipid metabolism, induced by ACSL5 over-expression, was characterized as crucial. ACSL5-related over-expression of mitochondrial mortalin was found in HEK293 and Lovo (wild-type TP53 [tumor protein p53]) and CaCo2 (p53-negative; TP53 mutated) cells but not in Colo320DM cells (mutated TP53). In normal human intestinal mucosa, an increasing gradient of both ACSL5 and mortalin from bottom to top was observed, whereas p53 (wild-type TP53) decreased. In sporadic intestinal adenocarcinomas with strong p53 immunostaining (mutated TP53), ACSL5-related mortalin expression was heterogeneous. ACSL5-induced mitochondrial mortalin expression is assumed to be a stress response to ACSL5-related changes in lipid metabolism and is regulated by the TP53 status. Uncoupling of ACSL5 and mitochondrial mortalin by mutated TP53 could be important in colorectal carcinogenesis.

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Acknowledgments

The authors are grateful to P. Akens for typing and proof-reading the manuscript. The great help of Dr. C. Henkel and S. Fiedler in preparation of protein gels and MALDI mass spectrometry and of D. Weisner in preparation of qRT-PCRs is highly appreciated.

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Authors and Affiliations

  1. Institute of Pathology, RWTH Aachen University, Pauwelsstrasse 30, 52074, Aachen, Germany

    Christina Klaus, Elke Kaemmerer, Andrea Reinartz, Ursula Schneider, Patrick Plum, Min Kyung Jeon, Josephine Hose, Franziska Hartmann & Nikolaus Gassler

  2. Department of Pediatrics, RWTH Aachen University, Aachen, Germany

    Elke Kaemmerer & Norbert Wagner

  3. Functional Proteome Analysis, DKFZ Heidelberg, Aachen, Germany

    Martina Schnölzer

  4. Institute of Pathology, Heidelberg University, Heidelberg, Germany

    Jürgen Kopitz

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  1. Christina Klaus
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  12. Nikolaus Gassler
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Correspondence to Nikolaus Gassler.

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Christina Klaus and Elke Kaemmerer contributed equally to this work.

None of the authors has any conflicts of interest.

This work was partially supported by the Deutsche Forschungsgemeinschaft (DFG GA 785/5-1) and Deutsche Krebshilfe (GA 109313).

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Klaus, C., Kaemmerer, E., Reinartz, A. et al. TP53 status regulates ACSL5-induced expression of mitochondrial mortalin in enterocytes and colorectal adenocarcinomas. Cell Tissue Res 357, 267–278 (2014). https://doi.org/10.1007/s00441-014-1826-8

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  • DOI: https://doi.org/10.1007/s00441-014-1826-8

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