The SARAH Domain of RASSF1A and Its Tumor Suppressor Function

Claudia Dittfeld,Antje Klagge-Ulonska,Antje M. Richter,Katrin Steinmann,Reinhard H. Dammann

doi:10.1155/2012/196715

Abstract

The Ras association domain family 1A (RASSF1A) tumor suppressor encodes a Sav-RASSF-Hpo domain (SARAH), which is an interaction domain characterized by hWW45 (dSAV) and MST1/2 (dHpo). In our study, the interaction between RASSF1A and RASSF1C with MST1 and MST2 was demonstrated and it was shown that this interaction depends on the SARAH domain. SARAH domain-deleted RASSF1A had a similar growth-reducing effect as full-length RASSF1A and inhibited anchorage independent growth of the lung cancer cell lines A549 significantly. In cancer cells expressing the SARAH deleted form of RASSF1A, reduced mitotic rates (P = 0.001) with abnormal metaphases (P < 0.001) were observed and a significantly increased rate of apoptosis was found (P = 0.006) compared to full-length RASSF1A. Although the association with microtubules and their stabilization was unaffected, mitotic spindle formation was altered by deletion of the SARAH domain of RASSF1A. In summary, our results suggest that the SARAH domain plays an important role in regulating the function of RASSF1A.

Highlights

The Ras association domain family 1 gene (RASSF1) was identified on chromosome 3p21.3, a region frequently deleted in cancer [1]
We demonstrate that the interaction of Ras association domain family 1A (RASSF1A) and RASSF1C with MST1 and MST2 depends on the Sav-RASSF-Hpo domain (SARAH) domain
To gain insight into the function of the SARAH domain of RASSF1, the interaction of RASSF1A, RASSF1C, RASSF1AΔSARAH, and RASSF1CΔSARAH with MST1 and MST2 was investigated in the yeast two-hybrid system (Figures 1(b) and 1(c))

Summary

Introduction

The Ras association domain family 1 gene (RASSF1) was identified on chromosome 3p21.3, a region frequently deleted in cancer [1]. The promoter of RASSF1A is often hypermethylated in cancer, whereas the promoter region of RASSF1C is never methylated [2, 3]. Both isoforms encode a Ras association domain in the C-terminus, an ATM-kinase phosphorylation site, a SARAH protein interaction domain, and the N-terminal sequence of RASSF1A harbors a diacyl glycerol binding domain [1, 4]. It has been reported that RASSF1A binds to microtubules and protects cells from microtubule destabilizing agents [7, 12,13,14,15] This interaction contributes to cell cycle regulation and mitotic progression

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