Molecular origin of the mosaic sequence arrangements of higher primate α-globin duplication units

The human adult α-globin locus consists of three pairs of homology blocks (X, Y, and Z) interspersed with three nonhomology blocks (I, II, and III), and three Alu family repeats, Alu1, Alu2, and Alu3. It has been suggested that an ancient primate α-globin-containing unit was ancestral to the X, Y, and Z and the Alu1/Alu2 repeats. However, the evolutionary origin of the three nonhomologous blocks has remained obscure. We have now analyzed the sequence organization of the entire adult α-globin locus of gibbon (Hylobates lar). DNA segments homologous to human block I occur in both duplication units of the gibbon α-globin locus. Detailed interspecies sequence comparisons suggest that nonhomologous blocks I and II, as well as another sequence, IV, were all part of the ancestral α-globin-containing unit prior to its tandem duplication. However, sometime thereafter, block I was deleted from the human α1-globin-containing unit, and block II was also deleted from the α2-globin-containing unit in both human and gibbon. These were probably independent events both mediated by independent illegitimate recombination processes. Interestingly, the end points of these deletions coincide with potential insertion sites of Alu family repeats. These results suggest that the shaping of DNA segments in eukaryotic genomes involved the retroposition of repetitive DNA elements in conjunction with simple DNA recombination processes.

CTXφ contains a hybrid genome derived from tandemly integrated elements

CTXφ is a filamentous, temperate bacteriophage whose genome includes ctxAB, the genes that encode cholera toxin. In toxigenic isolates of Vibrio cholerae, tandem arrays of prophage DNA, usually interspersed with the related genetic element RS1, are integrated site-specifically within the chromosome. We have discovered that these arrays routinely yield hybrid virions, composed of DNA from two adjacent prophages or from a prophage and a downstream RS1. Coding sequences are always derived from the 5′ prophage whereas most of an intergenic sequence, intergenic region 1, is always derived from the 3′ element. The presence of tandem elements is required for production of virions: V. cholerae strains that contain a solitary prophage rarely yield CTX virions, and the few virions detected result from imprecise excision of prophage DNA. Thus, generation of the replicative form of CTXφ, pCTX, a step that precedes production of virions, does not depend on reversal of the process for site-specific integration of CTXφ DNA into the V. cholerae chromosome. Production of pCTX also does not depend on RecA-mediated homologous recombination between adjacent prophages. We hypothesize that the CTXφ-specific proteins required for replication of pCTX can also function on a chromosomal substrate, and that, unlike the processes used by other integrating phages, production of pCTX and CTXφ does not require excision of the prophage from the chromosome. Use of this replication strategy maximizes vertical transmission of prophage DNA while still enabling dissemination of CTXφ to new hosts.

An integrated map of the genome of the tubercle bacillus, Mycobacterium tuberculosis H37Rv, and comparison with Mycobacterium leprae.

An integrated map of the genome of the tubercle bacillus, Mycobacterium tuberculosis, was constructed by using a twin-pronged approach. Pulsed-field gel electrophoretic analysis enabled cleavage sites for Asn I and Dra I to be positioned on the 4.4-Mb circular chromosome, while, in parallel, clones from two cosmid libraries were ordered into contigs by means of fingerprinting and hybridization mapping. The resultant contig map was readily correlated with the physical map of the genome via the landmarked restriction sites. Over 165 genes and markers were localized on the integrated map, thus enabling comparisons with the leprosy bacillus, Mycobacterium leprae, to be undertaken. Mycobacterial genomes appear to have evolved as mosaic structures since extended segments with conserved gene order and organization are interspersed with different flanking regions. Repetitive sequences and insertion elements are highly abundant in M. tuberculosis, but the distribution of IS6110 is apparently nonrandom.

Mg-SINE: a short interspersed nuclear element from the rice blast fungus, Magnaporthe grisea.

A short interspersed nuclear element, Mg-SINE, was isolated and characterized from the genome of the rice blast fungus, Magnaporthe grisea. Mg-SINE was isolated as an insertion element within Pot2, an inverted-repeat transposon from M. grisea and shows typical features of a mammalian SINE. Mg-SINE is present as a 0.47-kb interspersed sequence at approximately 100 copies per haploid genome in both rice and non-rice isolates of M. grisea, indicating a common evolutionary origin. Secondary structure analysis of Mg-SINE revealed a tRNA-related region at the 5' end which folds into a cloverleaf structure. Genomic fusions resulting in chimeric Mg-SINEs (Ch-SINEs) composed of a sequence homologous to Mg-SINE at the 3' end and an unrelated sequence at its 5' end were also isolated, indicating that this and other DNA rearrangements mediated by these elements may have a major effect on the genomic architecture of this fungus.

Repeated evolution of heat responsiveness among Brassicaceae COPIA transposable elements

Eukaryotic genomes contain repetitive DNA sequences. This includes simple repeats and more complex transposable elements (TEs). Many TEs reach high copy numbers in the host genome, owing to their amplification abilities by specific mechanisms. There is growing evidence that TEs contribute to gene transcriptional regulation. However, excess of TE activity may lead to reduced genome stability. Therefore, TEs are suppressed by the transcriptional gene silencing machinery via specific chromatin modifications. In contrary, effectiveness of the epigenetic silencing mechanisms imposes risk for TE survival in the host genome. Therefore, TEs may have evolved specific strategies for bypassing epigenetic control and allowing the emergence of new TE copies. Recent studies suggested that the epigenetic silencing can be, at least transiently, attenuated by heat stress in A. thaliana. Heat stress induced strong transcriptional activation of COPIA78 family LTR-retrotransposons named ONSEN, and even their transposition in mutants deficient in siRNA-biogenesis. ONSEN transcriptional activation was facilitated by the presence of heat responsive elements (HREs) within the long terminal repeats, which serve as a binding platform for the HEAT SHOCK FACTORs (HSFs). This thesis focused on the evolution of ONSEN heat responsiveness in Brassicaceae. By using whole-transcriptome sequencing approach, multiple Arabidopsis lyrata ONSENs with conserved heat response were found and together with ONSENs from other Brassicaceae were used to reconstruct the evolution of ONSEN HREs. This indicated ancestral situation with two, in palindrome organized, HSF binding motifs. In the genera Arabidopsis and Ballantinia, a local duplication of this locus increased number of HSF binding motifs to four, forming a high-efficiency HRE. In addition, whole transcriptome analysis revealed novel heat-responsive TE families COPIA20, COPIA37 and HATE. Notably, HATE represents so far unknown COPIA family which occurs in several Brassicaceae species but is absent in A. thaliana. Putative HREs were identified within the LTRs of COPIA20, COPIA37 and HATE of A. lyrata, and could be preliminarily validated by transcriptional analysis upon heat induction in subsequent survey of Brassicaeae species. Subsequent phylogenetic analysis indicated a repeated evolution of heat responsiveness within Brassicaceae COPIA LTR-retrotransposons. This indicates that acquisition of heat responsiveness may represent a successful strategy for survival of TEs within the host genome.

Case definition and holistic model modifications required for building elements

This project is an extension of a previous CRC project (220-059-B) which developed a program for life prediction of gutters in Queensland schools. A number of sources of information on service life of metallic building components were formed into databases linked to a Case-Based Reasoning Engine which extracted relevant cases from each source.

Report : holistic model modifications for selected building elements

This project is an extension of a previous CRC project (220-059-B) which developed a program for life prediction of gutters in Queensland schools. A number of sources of information on service life of metallic building components were formed into databases linked to a Case-Based Reasoning Engine which extracted relevant cases from each source.

Selected elements of housing affordability impacting otherwise potentially sustainable communities

It is widely held that strong relationships exist between housing, economic status, and well being. Therefore, recent events emerging from the United States, culminating in widespread housing stock surpluses in that country and others, threaten to destabilise many aspects related to individuals and community. However, despite global impact, the position of housing demand and supply is not consistent. The Australian position provides a strong contrast whereby continued strong housing demand generally remains a critical issue affecting the socio-economic landscape. Underpinned by strong levels of immigration, and further buoyed by sustained historically low interest rates, increasing income levels, and increased government assistance for first home buyers, this strong housing demand ensures elements related to housing affordability continue to gain prominence. A significant, but less visible factor impacting housing affordability – particularly new housing development – relates to holding costs. These costs are in many ways “hidden” and cannot always be easily identified. Although it is only one contributor, the nature and extent of its impact requires elucidation. In its simplest form, it commences with a calculation of the interest or opportunity cost of land holding. However, there is significantly more complexity for major new developments - particularly greenfield development. Analysis suggests that even small shifts in primary factors impacting holding costs can appreciably affect housing affordability. Those factors of greatest significance not only include interest rates and the rate of inflation, but even less apparent factors such as the regulatory assessment period. These are not just theoretical concepts but real, measurable price drivers. Ultimately, the real impact is felt by the one market segment whom can typically least afford it – new home, first home buyers. They can be easily pushed out of affordability. This paper suggests the stability and sustainability of growing, new communities require this problem to be acknowledged and accurately identified if the well being of such communities is to be achieved.

A numerical method to quantify the axial shortening of vertical elements in concrete buildings

Differential axial shortening, distortion and deformation in high rise buildings is a serious concern. They are caused by three time dependent modes of volume change; “shrinkage”, “creep” and “elastic shortening” that takes place in every concrete element during and after construction. Vertical concrete components in a high rise building are sized and designed based on their strength demand to carry gravity and lateral loads. Therefore, columns and walls are sized, shaped and reinforced differently with varying concrete grades and volume to surface area ratios. These structural components may be subjected to the detrimental effects of differential axial shortening that escalates with increasing the height of buildings. This can have an adverse impact on other structural and non-structural elements. Limited procedures are available to quantify axial shortening, and the results obtained from them differ because each procedure is based on various assumptions and limited to few parameters. All these prompt to a need to develop an accurate numerical procedure to quantify the axial shortening of concrete buildings taking into account the important time varying functions of (i) construction sequence (ii) Young’s Modulus and (iii) creep and shrinkage models associated with reinforced concrete. General assumptions are refined to minimize variability of creep and shrinkage parameters to improve accuracy of the results. Finite element techniques are used in the procedure that employs time history analysis along with compression only elements to simulate staged construction behaviour. This paper presents such a procedure and illustrates it through an example. Keywords: Differential Axial Shortening, Concrete Buildings, Creep and Shrinkage, Construction Sequence, Finite Element Method.