Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

NHERF (Na+/H+ Exchanger Regulatory Factor) gene mutations in human breast cancer

Abstract

Yeast two-hybrid screening was used to explore novel proteins that interact with a breast tumor or metastasis suppressor, SYK (spleen tyrosine kinase). The screening yielded NHERF (Na+/H+ exchanger regulatory factor, also known as NHERF1 or EBP-50) that binds to the interdomain B of SYK. NHERF is an estrogen-responsive gene that encodes an inhibitory factor for epithelial Na+/H+ exchanger isoform 3 (NHE3). We found intragenic mutation of the NHERF gene accompanied by loss of heterzygosity (LOH) in 3% (3/85) of breast cancer cell lines and primary breast tumors. Mutations occurred at the conserved PDZ domains at NHERF NH2-terminus that bound to SYK, or at its COOH-terminus motif that binds to MERLIN, the product of Neurofibromatosis 2 (NF2) tumor suppressor gene. NHERF tumorigenic mutations decreased or abolished its interaction with SYK or MERLIN, suggesting a pathway link among these three molecules that may play a critical role in mammary neoplastic progression. Primary breast tumors with LOH at the NHERF locus had clinical presentations of higher aggressiveness, indicating that deregulated NHERF signaling may be associated with disease progression. Moreover, the LOH was inversely correlated with SYK promoter methylation, suggesting that NHERF and SYK may transduce a common suppressive signal. Taken together, the results indicated NHERF to be a candidate tumor suppressor gene in human breast carcinoma that may be interconnected to the SYK and MERLIN suppressors.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4

Similar content being viewed by others

Abbreviations

LOH:

loss of heterozygosity

PCR:

polymerase chain reaction

RT–PCR:

reverse transcriptase–PCR

SYK:

spleen tyrosine kinase

IDB:

interdomain B

NHERF:

Na+/H+ exchanger regulatory factor

NHE3:

Na+/H+ exchanger isoform 3

NF2:

neurofibromatosis 2

IP:

immunoprecipitation

IB:

immunoblotting

GST:

glutathione-S-transferase

IPTG:

isopropyl β-D-1-thiogalactopyranoside

β-gal:

β-galactosidase

ER:

estrogen receptor

PR:

progesterone receptor

PDZD:

PDZ domain

References

  • Arakawa H, Hayashi N, Nagase H, Ogawa M and Nakamura Y . (1994). Hum. Mol. Genet., 3, 565–568.

  • Bartkova J, Lukas J, Guldberg P, Alsner J, Kirkin AF, Zeuthen J and Bartek J . (1996). Cancer Res., 56, 5475–5483.

  • Beckmann MW, Niederacher D, Schnurch HG, Gusterson BA and Bender HG . (1997). J. Mol. Med., 75, 429–439.

  • Bianchi AB, Hara T, Ramesh V, Gao J, Klein-Szanto AJ, Morin F, Menon AG, Trofatter JA, Gusella JF, Seizinger BR and Kley N . (1994). Nat. Genet., 6, 185–192.

  • Coopman PJ, Do MT, Barth M, Bowden ET, Hayes AJ, Basyuk E, Blancato JK, Vezza PR, McLeskey SW, Mangeat PH and Mueller SC . (2000). Nature, 406, 742–747.

  • Couch FJ and Weber BL . (2002). The Genetic Basis of Human Cancer Vogesltein B & Kinzler KW (eds). McGraw-Hill Companies Inc.: New York, pp. 549–581.

    Google Scholar 

  • Ediger TR, Kraus WL, Weinman EJ and Katzenellenbogen BS . (1999). Endocrinology, 140, 2976–2982.

  • Hall RA, Premont RT, Chow CW, Blitzer JT, Pitcher JA, Claing A, Stoffel RH, Barak LS, Shenolikar S, Weinman EJ, Grinstein S and Lefkowitz RJ . (1998). Nature, 392, 626–630.

  • Harkes IC, Elstrodt F, Dinjens WN, Molier M, Klijn JG, Berns EM and Schutte M . (2003). Br. J. Cancer, 89, 2289–2292.

  • Harris AL . (2002). Nat. Rev., 2, 38–47.

  • Kalikin LM, Frank TS, Svoboda-Newman SM, Wetzel JC, Cooney KA and Petty EM . (1997). Oncogene, 14, 1991–1994.

  • Kalikin LM, Qu X, Frank TS, Caduff RF, Svoboda SM, Law DJ and Petty EM . (1996). Genes Chromosome Cancer, 17, 64–68.

  • Kucherlapati M, Yang K, Kuraguchi M, Zhao J, Lia M, Heyer J, Kane MF, Fan K, Russell R, Brown AM, Kneitz B, Edelmann W, Kolodner RD, Lipkin M and Kucherlapati R . (2002). PNAS, 99, 9924–9929.

  • Lupulescu A . (1993). Exp. Clin. Endocrinol., 101, 204–214.

  • Magee JA, Abdulkadir SA and Milbrandt J . (2003). Cancer Cell, 3, 273–283.

  • Mahabeleshwar GH and Kundu GC . (2003). J. Biol. Chem., 278, 6209–6221.

  • Mohler PJ, Kreda SM, Boucher RC, Sudol M, Stutts MJ and Milgram SL . (1999). J. Cell Biol., 147, 879–890.

  • Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B and Kinzler KW . (1997). Science, 275, 1787–1790.

  • Murthy A, Gonzalez-Agosti C, Cordero E, Pinney D, Candia C, Solomon F, Gusella J and Ramesh V . (1998). J. Biol. Chem., 273, 1273–1276.

  • Nandi S, Guzman R and Yang J . (1995). PNAS, 92, 3650–3657.

  • Orsetti B, Courjal F, Cuny M, Rodriguez C and Theillet C . (1999). Oncogene, 18, 6262–6270.

  • Plummer SJ, Paris MJ, Myles J, Tubbs R, Crowe J and Casey G . (1997). Genes Chromosome Cancer, 20, 354–362.

  • Radford DM, Fair KL, Philips NJ, Ritter JH, Steinbrueck T, Holt MS and Donis-Keller H . (1995). Cancer Res., 55, 3399–3405.

  • Reczek D, Berryman M and Bretscher A . (1997). J. Cell Biol., 139, 169–179.

  • Reifenberger G, Liu L, Ichimura K, Schmidt EE and Collins VP . (1993). Cancer Res., 53, 2736–2739.

  • Rouleau GA, Merel P, Lutchman M, Sanson M, Zucman J, Marineau C, Hoang-Xuan K, Demczuk S, Desmaze C, Plougastel B, Pulst SM, Lenoir G, Bijlsma E, Fashold R, Dumanski J, de Jong P, Parry D, Eldrige R, Aurias A, Delattre O and Thomas G . (1993). Nature, 363, 515–521.

  • Srivastava M, Montagna C, Leighton X, Glasman M, Naga S, Eidelman O, Ried T and Pollard HB . (2003). PNAS, 100, 14287–14292.

  • Staff S, Isola J and Tanner M . (2003). Cancer Res., 63, 4978–4983.

  • Stemmer-Rachamimov AO, Wiederhold T, Nielsen GP, James M, Pinney-Michalowski D, Roy JE, Cohen WA, Ramesh V and Louis DN . (2001). Am. J. Pathol., 158, 57–62.

  • Stubbs M, McSheehy PMJ, Griffiths JR and Bashford CL . (2000). Mol. Med. Today, 6, 15–19.

  • Toyama T, Iwase H, Yamashita H, Hara Y, Omoto Y, Sugiura H, Zhang Z and Fujii Y . (2002). Cancer Lett., 189, 97–102.

  • Trofatter JA, MacCollin MM, Rutter JL, Murrell JR, Duyao MP, Parry DM, Eldridge R, Kley N, Menon AG, Pulaski K, Haase VH, Ambrose CM, Munroe D, Bove C, Haines JL, Martuza RL, MacDonald ME, Seizinger BR, Short MP, Buckler AJ and Gusella JF . (1993). Cell, 72, 791–800.

  • Wang L, Duke L, Zhang PS, Arlinghaus RB, Symmans WF, Sahin A, Mendez R and Dai JL . (2003). Cancer Res., 63, 4724–4730.

  • Wang S, Raab RW, Schatz PJ, Guggino WB and Li M . (1998). FEBS Lett., 427, 103–108.

  • Weinman EJ, Steplock D, Donowitz M and Shenolikar S . (2000). Biochemistry, 39, 6123–6129.

  • Weinman EJ, Steplock D and Shenolikar S . (2003). FEBS Lett., 2003, 141–144.

  • Yaegashi S, Sachse R, Ohuchi N, Mori S and Sekiya T . (1995). Jpn. J. Cancer Res., 86, 923–933.

  • Yuan YF, Mendez R, Sahin A and Dai JL . (2001). Cancer Res., 61, 5558–5561.

Download references

Acknowledgements

We thank Drs N DeVleeschouwer, S Ethier, H Smith, and E Stockert for sharing genomic DNAs and cell lines, Dr PS Zhang for technical help, and Ms L Feldman and CY Wang for editing. This work was supported by an R01 grant (CA100278) from the National Institutes of Health (to JLD).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jia Le Dai.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Dai, J., Wang, L., Sahin, A. et al. NHERF (Na+/H+ Exchanger Regulatory Factor) gene mutations in human breast cancer. Oncogene 23, 8681–8687 (2004). https://doi.org/10.1038/sj.onc.1207962

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1207962

Keywords

This article is cited by

Search

Quick links