Universal two-point interaction of mediator KIX with 9aaTAD activation domains.

The nine-amino-acid activation domain (9aaTAD) is defined by a short amino acid pattern including two hydrophobic regions (positions p3-4 and p6-7). The KIX domain of mediator transcription CBP interacts with the 9aaTAD domains of transcription factors MLL, E2A, NF-kB, and p53. In this study, we analyzed the 9aaTADs-KIX interactions by nuclear magnetic resonance. The positions of three KIX helixes α1-α2-α3 are influenced by sterically-associated hydrophobic I611, L628, and I660 residues that are exposed to solvent. The positions of two rigid KIX helixes α1 and α2 generate conditions for structural folding in the flexible KIX-L12-G2 regions localized between them. The three KIX I611, L628, and I660 residues interact with two 9aaTAD hydrophobic residues in positions p3 and p4 and together build a hydrophobic core of five residues (5R). Numerous residues in 9aaTAD position p3 and p4 could provide this interaction. Following binding of the 9aaTAD to KIX, the hydrophobic I611, L628, and I660 residues are no longer exposed to solvent and their position changes inside the hydrophobic core together with position of KIX α1-α2-α3 helixes. The new positions of the KIX helixes α1 and α2 allow the KIX-L12-G2 enhanced formation. The second hydrophobic region of the 9aaTAD (positions p6 and p7) provides strong binding with the KIX-L12-G2 region. Similarly, multiple residues in 9aaTAD position p6 and p7 could provide this interaction. In conclusion, both 9aaTAD regions p3, p4 and p6, p7 provide co-operative and highly universal binding to mediator KIX. The hydrophobic core 5R formation allows new positions of the rigid KIX α-helixes and enables the enhanced formation of the KIX-L12-G2 region. This contributes to free energy and is the key for the KIX-9aaTAD binding. Therefore, the 9aaTAD-KIX interactions do not operate under the rigid key-and-lock mechanism what explains the 9aaTAD natural variability.

Prospective Multiparametric Magnetic Resonance Monitoring of Changes in Lesions of Hyaline Cartilage of the Knee Joint After Treatment by Microfractures and Implantation of Biological Collagen Type I Matrix Implants .

RATIONALE AND OBJECTIVES: This study's aims were to depict changes in cartilage quality after surgical intervention using magnetic resonance (MR) examination and in content of glycosaminoglycans chains (GAGs) after two types of surgeries - chondral defect treatment by microfractures and scaffold implantation in combination with microfractures. MATERIALS AND METHODS: Twenty-five patients were studied: 14 with implants, 11 with microfractures. MR examination was made before surgery and 6, 12, and 18 months thereafter. Qualitative changes in cartilage were observed by means of delayed gadolinium enhanced magnetic resonance imaging of cartilage sequence using Gd-DTPA2- and Gd-DOTA. In each examination, GAGs content was determined at three locations: the defect, its surroundings, and a non-load-bearing reference area. RESULTS: Measured indices showed no statistically significant differences in changes within the defect area when comparing the two treatment types at individual time points of 6, 12, and 18 months. In the case of microfracture treatment, more substantial decrease in GAGs concentration occurred at month 6, whereas the greatest decline occurred at month 12 when using an implant. Change in GAGs content and decline in cartilage quality were substantial also in the reference area and close surroundings. CONCLUSIONS: Hyaline cartilage behaves as a unified whole, and change in GAGs content was marked also in locations with no morphological damage. Over the monitored period, no statistically significant difference between treatment types was noted as measured by GAGs content in the defect or its close surroundings. dGEMRIC is suitable for monitoring cartilage quality even if use of Gd-DTPA2- is not possible, because comparable results were achieved using Gd-DOTA.