Mapping of the critical region of mitogene-inducible gene-6 for NF-kappaB activation.

Abstract:

:Mitogene-inducible gene-6 (Mig-6) is involved in the intracellular signaling pathway as an adaptor molecule. Mig-6 expression is rapidly induced upon various kinds of mitogenic and stressful stimulations. We previously demonstrated that Mig-6 expression is induced by activated Ki-ras in human colon cancer cells and the limited proteolytic processed NH(2)-terminal region containing the Cdc42/Rac interaction and binding (CRIB) domain of Mig-6 is bound to IkappaBalpha, resulting in NF-kappaB activation. In this study, we tried to determine the critical region of Mig-6 for regulating NF-kappaB activation by using various kinds of deletion-constructs of Mig-6. The CRIB domain-deleted Mig-6 fragment (residues 68 to 462) did not show significant NF-kappaB activation. Furthermore, NH(2)-terminal 148, 158 and 180 amino acid regions of Mig-6 did not show NF-kappaB activation. On the other hand, the NH(2)-terminal 264 amino acid region of Mig-6 did show NF-kappaB activation to a similar extent of the full length of Mig-6. Interestingly, the full length and NH(2)-terminal 264 amino acid region of Mig-6 produced two kinds of cleaved NH(2)-terminal fragments, N1 and N2 with an approximate molecular size of 5 and 7 kDa, respectively. The NH(2)-terminal 148 and 158 amino acid regions of Mig-6 did not produce N1 or N2 fragments, while the NH(2)-terminal 180 amino acid region of Mig-6 only produced the N1 fragment. Furthermore, the N2 fragment was bound with IkappaBalpha, but the N1 fragment was not. These results together suggested that the NH(2)-terminal 264 amino acid region is critical for NF-kappaB activation and proper limited proteolytic processing of Mig-6.

journal_name

Oncol Rep

journal_title

Oncology reports

authors

Mabuchi R,Sasazuki T,Shirasawa S

keywords:

subject

Has Abstract

pub_date

2005-03-01 00:00:00

pages

473-6

issue

3

eissn

1021-335X

issn

1791-2431

journal_volume

13

pub_type

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