ABCMdb

PMID: 26173179

Ito A, Hong C, Rong X, Zhu X, Tarling EJ, Hedde PN, Gratton E, Parks J, Tontonoz P
LXRs link metabolism to inflammation through Abca1-dependent regulation of membrane composition and TLR signaling.
Elife. 2015 Jul 14;4:e08009. doi: 10.7554/eLife.08009., [PubMed]

Sentences

No. Mutations Sentence Comment

46 ABCA1 p.Leu439Ala (C) Immortalized MEFs from Lxrb1;-/-Lxrb2; -/- mice reconstituted with wild-type human LXRb1;, L439A/E441A mutant or control mock were pretreated with the LXR agonist GW3965 (1 bc;M) overnight, followed by stimulation with LPS (10 ng/ml) for 4 hr. Gene expression was analyzed by real-time PCR. Login to comment 54 ABCA1 p.Leu439Ala Furthermore, an LXR mutant defective in its ability to recruit co-activators, L439A/E441A (Tzukerman et al., 1994; Bastie et al., 2000), was unable to induce Abca1 or to repress inflammatory genes in both MEFs and iBMDM (Figure 1C, Figure 2B, Figure 2-figure supplement 1A), strongly suggesting that gene activation and inflammatory repression are mechanistically linked. Login to comment 80 ABCA1 p.Leu439Ala (B, C) Immortalized bone marrow-derived macrophages from Lxrb1;-/-Lxrb2;-/- mice reconstituted with wild-type human LXRb1;, K328R/K434R (KK) mutant, L439A/E441A mutant, or mock control were pretreated with GW3965 (1 bc;M) overnight, followed by stimulation with LPS (10 ng/ml) for 4 hr. Gene expression was analyzed by real-time PCR (B) and Agilent microarrays (C). Login to comment 81 ABCA1 p.Cys1477Arg ABCA1 p.Asn935Ser Selected genes that are annotated with the 'Immune system process` GO Figure 2. continued on next page To further examine the function of Abca1 was critical for inflammatory repression, we reconstituted iBMDM from myeloid-specific Abca1-/- mice with wild-type Abca1 or two different Abca1 point mutants that lacks cholesterol efflux ability, N935S and C1477R (Singaraja et al., 2006; Kannenberg et al., 2013). Login to comment 82 ABCA1 p.Cys1477Arg ABCA1 p.Asn935Ser When Abca1-deficient cells were reconstituted with wild-type Abca1, inflammatory gene expression was repressed by LXR activation; however, this effect was lost in cells expressing the N935S or C1477R mutants (Figure 4C). Login to comment 149 ABCA1 p.Asn935Ser (C) Immortalized bone marrow-derived macrophages from Abca1-/- mice reconstituted with wild-type Abca1, N935S mutant, C1447R mutant or mock control were pretreated with GW3965 (1 bc;M) overnight, followed by stimulation with LPS (10 ng/ml) for 4 hr. Gene expression was analyzed by real-time PCR. Login to comment 184 ABCA1 p.Leu439Ala We found that an LXR L439A/E441A mutant that lacks co-activator recruitment capacity was unable to repress inflammation. Login to comment