Specific binding of the mammalian high mobility group protein AT-hook 2 to the minor groove of AT -rich DNAs: Thermodynamic and specificity studies

The mammalian high mobility group protein AT-hook 2 (HMGA2) is a small transcriptional factor involved in cell development and oncogenesis. It contains three "AT-hook" DNA binding domains, which specifically recognize the minor groove of AT-rich DNA sequences. It also has an acidic C-terminal motif. Previous studies showed that HMGA2 mediates all its biological effects through interactions with AT-rich DNA sequences in the promoter regions. In this dissertation, I used a variety of biochemical and biophysical methods to examine the physical properties of HMGA2 and to further investigate HMGA2's interactions with AT-rich DNA sequences. The following are three avenues perused in this study: (1) due to the asymmetrical charge distribution of HMGA2, I have developed a rapid procedure to purify HMGA2 in the milligram range. Preparation of large amounts of HMGA2 makes biophysical studies possible; (2) Since HMGA2 binds to different AT-rich sequences in the promoter regions, I used a combination of isothermal titration calorimetry (ITC) and DNA UV melting experiment to characterize interactions of HMGA2 with poly(dA-dT) 2 and poly(dA)poly(dT). My results demonstrated that (i) each HMGA2 molecule binds to 15 AT bp; (ii) HMGA2 binds to both AT DNAs with very high affinity. However, the binding reaction of HMGA2 to poly(dA-dT) 2 is enthalpy-driven and the binding reaction of HMGA2 with poly(dA)poly(dT) is entropy-driven; (iii) the binding reactions are strongly depended on salt concentrations; (3) Previous studies showed that HMGA2 may have sequence specificity. In this study, I used a PCR-based SELEX procedure to examine the DNA binding specificity of HMGA2. Two consensus sequences for HMGA2 have been identified: 5'-ATATTCGCGAWWATT-3' and 5'-ATATTGCGCAWWATT-3', where W represents A or T. These consensus sequences have a unique feature: the first five base pairs are AT-rich, the middle four to five base pairs are GC-rich, and the last five to six base pairs are AT-rich. All three segments are critical for high affinity binding. Replacing either one of the AT-rich sequences to a non-AT-rich sequence causes at least 100-fold decrease in the binding affinity. Intriguingly, if the GC-segment is substituted by an AT-rich segment, the binding affinity of HMGA2 is reduced approximately 5-fold. Identification of the consensus sequences for HMGA2 represents an important step towards finding its binding sites within the genome.

Social support, risk, and adjustment of immigrant preadolescents

The current study was designed to explore the salience of social support, immigrant status, and risk in middle childhood and early adolescence across two time periods as indicated by measures of school adjustment and well-being. Participants included 691 children of public elementary schools in grades 4 and 6 who were interviewed in 1997 (Time 1) and reinterviewed two years later (Time 2); 539 were U.S.-born, and 152 were foreign-born. ^ Repeated measures multivariate analyses of variance (MANOVA's) were conducted to assess the effects of immigrant status and risk on total support, well-being, and school adjustment from Time 1 to Time 2. Follow-up analyses, including Student-Newman-Keuls post hoc tests, were used to test the significance of the differences among the means of support categories (low and high), immigrant status (U.S. born and non-U.S. born), risk (low and high) and time (time 1 and time 2). ^ Results showed that immigrant participants in the high risk group reported significantly lower levels of support than their peers. Further, children of low risk at Time 2 indicated the highest levels of support. Second, immigrant preadolescents, preadolescents who reported low levels of social support, and preadolescents of the high risk reported lower levels of emotional well-being. There was also an interaction of support by risk by time, indicating that children who are at risk and had low levels of social support reported more emotional problems at Time 1. Finally, preadolescents who are at risk and preadolescents who reported lower levels of support were more likely to show school adaptation problems. Findings from this study highlight the importance of a multivariable approach to the study of support, emotional adjustment, and academic adjustment of immigrant preadolescents. ^

Orthogonal frequency division multiplexing based air interfaces and multiple input multiple output techniques in cooperative satellite communications for 4th generation mobile systems

Recently, wireless network technology has grown at such a pace that scientific research has become a practical reality in a very short time span. Mobile wireless communications have witnessed the adoption of several generations, each of them complementing and improving the former. One mobile system that features high data rates and open network architecture is 4G. Currently, the research community and industry, in the field of wireless networks, are working on possible choices for solutions in the 4G system. 4G is a collection of technologies and standards that will allow a range of ubiquitous computing and wireless communication architectures. The researcher considers one of the most important characteristics of future 4G mobile systems the ability to guarantee reliable communications from 100 Mbps, in high mobility links, to as high as 1 Gbps for low mobility users, in addition to high efficiency in the spectrum usage. On mobile wireless communications networks, one important factor is the coverage of large geographical areas. In 4G systems, a hybrid satellite/terrestrial network is crucial to providing users with coverage wherever needed. Subscribers thus require a reliable satellite link to access their services when they are in remote locations, where a terrestrial infrastructure is unavailable. Thus, they must rely upon satellite coverage. Good modulation and access technique are also required in order to transmit high data rates over satellite links to mobile users. This technique must adapt to the characteristics of the satellite channel and also be efficient in the use of allocated bandwidth. Satellite links are fading channels, when used by mobile users. Some measures designed to approach these fading environments make use of: (1) spatial diversity (two receive antenna configuration); (2) time diversity (channel interleaver/spreading techniques); and (3) upper layer FEC. The author proposes the use of OFDM (Orthogonal Frequency Multiple Access) for the satellite link by increasing the time diversity. This technique will allow for an increase of the data rate, as primarily required by multimedia applications, and will also optimally use the available bandwidth. In addition, this dissertation approaches the use of Cooperative Satellite Communications for hybrid satellite/terrestrial networks. By using this technique, the satellite coverage can be extended to areas where there is no direct link to the satellite. For this purpose, a good channel model is necessary.