The rapamycin-binding domain governs substrate selectivity by the mammalian target of rapamycin.

Abstract:

:The mammalian target of rapamycin (mTOR) is a Ser/Thr (S/T) protein kinase, which controls mRNA translation initiation by modulating phosphorylation of the translational regulators PHAS-I and p70(S6K). Here we show that in vitro mTOR is able to phosphorylate these two regulators at comparable rates. Both (S/T)P sites, such as Thr36, Thr45, and Thr69 in PHAS-I and the h(S/T)h site (where h is a hydrophobic amino acid) Thr389 in p70(S6K), were phosphorylated. Rapamycin-FKBP12 inhibited mTOR activity. Surprisingly, the extent of inhibition depended on the substrate. Moreover, mutating Ser2035 in the rapamycin-binding domain (FRB) not only decreased rapamycin sensitivity as expected but also dramatically affected the sites phosphorylated by mTOR. The results demonstrate that mutations in Ser2035 are not silent with respect to mTOR activity and implicate the FRB in substrate recognition. The findings also impose new limitations on interpreting results from experiments in which rapamycin and/or rapamycin-resistant forms of mTOR are used to investigate mTOR function in cells.

journal_name

Mol Cell Biol

authors

McMahon LP,Choi KM,Lin TA,Abraham RT,Lawrence JC Jr

doi

10.1128/mcb.22.21.7428-7438.2002

subject

Has Abstract

pub_date

2002-11-01 00:00:00

pages

7428-38

issue

21

eissn

0270-7306

issn

1098-5549

journal_volume

22

pub_type

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