PMC

SAXS experimental data, Kratky plots, and P(r)s for two dENE constructs and their complexes: wild-type dENE (A) and Xtal dENE (C) constructs. A.U., arbitrary units. The reconstructions of the wild-type, Xtal dENE constructs and their poly(A)₃₀ complexes are shown in B and D (cross-sections on the Right). (Scale bars: 20 Å.) The color coding presented in A and C apply to B and D as well. The histograms compare the obtained values for R_g (E) and D_max (F) for the wild-type and Xtal dENE constructs. The error bars in E were obtained directly from the Guinier analysis. The error bars in F were computed by sampling 25 values close to D_max, with individual weights based on the metric α ().

“Sharing Bioethics” shows dozens of land-based teachings. At first glance, the viewer can identify two intersecting circles of medicine. A circle surrounds a clinician in a white coat and bioscientific tools of medicine, such as a stethoscope and x-rays. In the Dene circle of medicine, plant medicine and Dene medicine stories are represented by a medicine bundle, strawberries and antlers. In the space where the circles intersect, the two healers also connect in a new space of relationship and possibility. In this new circle of medicine, neither knowledge or perspective is lost.

Cryo-EM structure of the B dENE construct without poly(A) at 8.7-Å resolution. (A and B) Density maps displayed at two contour levels (2.5σ, A; 3.5σ, B). The arrow indicates a hole in the lower dENE domain. (C) The dENE+poly(A) complex cryo-EM map with 8.7-Å resolution low-pass filtering, with poly(A) density subtracted (contoured at 3σ, salmon) and (D) its alignment with the free dENE map (contoured at 3.5σ, green). The two holes in C are in the upper and the lower URILs that form major-groove triple helices with poly(A). Upon poly(A) binding, local conformational changes occur in the lower dENE domain.

The TWIFB1 dENE pocket motif blocks the poly(A) exit path, forming a 3ʹ blunt-ended triplex. (A) Molecular surface representations of different ENEs oriented to show the direction of poly(A)/A-rich tract (white open arrow), the 3ʹ-most adenosine of each major-groove triplex and its surrounding region. (B) Superposition of the triple helices from various ENEs using the last four U-A•U triples and three base pairs of their flanking stems. The upper dENE domain is superimposed over the PAN or MALAT1 ENE with an RMSD = 1.1 or 1.2 Å, respectively. In the superposition of the two dENE domains, only the major-groove triple helices were used (RMSD = 0.7 Å). The upper dENE triplex on right is shown in transparent colors. The red-colored region represents predicted steric clash that should occur between the backbone of poly(A) and the pocket motif if the poly(A) strand leaves the major-groove triplex in the lower dENE domain. PAN and MALAT1 triplex structures were derived from PDB 3P22 () and 4PLX (), respectively. (C) Cartoon representation of the TWIFB1 dENE with a 72-mer poly(A) tail. Us changed to Cs in M26 are circled in orange. (D) In vitro deadenylation assays show that wild-type dENE protects its downstream poly(A) from deadenylation, while the absence of the pocket in M11 abrogates this ability. In M26, part of the poly(A) tail is trimmed rapidly and a product of the same size as wild-type dENE-poly(A)₇₂ accumulates. dT lanes show transcripts fully deadenylated by endogenous RNase H in the HeLa cell nuclear extract upon addition of oligo(dT)₄₀ to the reaction.