The putative tumor suppressor RASSF1A homodimerizes and heterodimerizes with the Ras-GTP binding protein Nore1.
Nore and RASSF1A are noncatalytic proteins that share 50% identity over their carboxyterminal 300 AA, a segment that encompasses a putative Ras-Rap association (RA) domain. RASSF1 is expressed as several splice variants, each of which contain an RA domain, however the 340 AA RASSF1A, but not the shorter RASSF1C variant, is a putative tumor suppressor. Nore binds to Ras and several Ras-like GTPases in a GTP dependent fashion however neither RASSF1 (A or C) or the C. elegans Nore/RASSF1 homolog, T24F1.3 exhibit any interaction with Ras or six other Ras-like GTPases in a yeast two-hybrid expression assay. A low recovery of RASSF1A (but not RASSF1C) in association with RasG12V is observed however on transient expression in COS cells. Nore and RASSF1A can each efficiently homodimerize and heterodimerize with each other through their nonhomologous aminoterminal segments. Recombinant RASSF1C exhibits a much weaker ability to homodimerize or heterodimerize; thus the binding of RASSF1C to Nore is very much less than the binding of RASSF1A to Nore. The association of RASSF1A with RasG12V in COS cells appears to reflect the heterodimerization of RASSF1A with Nore, inasmuch the recovery of RASSF1A with RasG12V is increased by concurrent expression of full length Nore, and abolished by expression of Nore deleted of its RA domain. The preferential ability of RASSF1A to heterodimerize with Nore and thereby associate with Ras-like GTPases may be relevant to its putative tumor suppressor function.
Ortiz-Vega
Sara
S
Diabetes Unit and Medical Services, Massachusetts General Hospital, Boston, Massachusetts, MA 02114, USA.
Khokhlatchev
Andrei
A
Nedwidek
Maria
M
Zhang
Xian-feng
XF
Dammann
Reinhard
R
Pfeifer
Gerd P
GP
Avruch
Joseph
J
D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D029742:Caenorhabditis elegans Proteins; D019281:Dimerization; D016147:Genes, Tumor Suppressor; D008970:Molecular Weight; D020559:Monomeric GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D025521:Tumor Suppressor Proteins; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D029742:Caenorhabditis elegans Proteins; D009363:Neoplasm Proteins; C411762:RASSF1 protein, human; D012333:RNA, Messenger; C578982:RSF-1 protein, C elegans; D025521:Tumor Suppressor Proteins; D020559:Monomeric GTP-Binding Proteins