Localisation of MT1-MMP to motility-associated structures

MT1-MMP is a crucial enzyme for cellular invasion in tissues, and its polarised cell surface localisation is thought to be an important regulatory mechanism. In this thesis, the mechanisms regulating localisation of MT1-MMP to motility-associated structures were investigated. Using a series of MT1-MMP domain deletion mutants, it was found that an eight amino acid region in the catalytic domain called the “MT-Loop” is involved in localisation of the enzyme to matrix attachment sites. A previously described mechanism by which MT1-MMP localises to lamellipodia is association with CD44. Interestingly, knockdown of the CD44 in HT-1080 cells increased MT1-MMP localisation to the matrix attachment sites, whereas CD44 overexpression in COS-7 cells decreased this localisation. These results indicate that CD44-mediated localisation to lamellipodia may compete with the localisation to the matrix attachment sites. One of the previously reported mechanisms localising MT1-MMP to the matrix attachment sites is by targeting the enzyme to cortactin-mediated invadopodia structures. However, silencing of the cortactin gene in HT-1080 cells unexpectedly enhanced this localisation, suggesting that invadopodia may not be required. For MT1-MMP to appear on the cell surface, the newly expressed molecules need to be trafficked from the trans-Golgi network to the plasma membrane by kinesin motor proteins (KIFs). 17 KIFs were tested for their involvement in the intracellular trafficking of MT1-MMP-containing vesicles. Silencing of the KIF3A or KIF13A genes markedly decreased MT1-MMP-dependent gelatin and collagen film degradation, whereas silencing of the KIF9 and KIF1C genes enhanced degradation of the underlying matrix. These and other data presented in this thesis suggest that KIF3A, KIF13A, KIF9 and KIF1C may be involved in vesicle trafficking of MT1-MMP to different areas of the plasma membrane and regulate spatial localisation of the enzyme. Taken together, these findings indicate that polarised cell surface localisation of MT1-MMP is regulated by multiple mechanisms, which may be important for the enzyme to promote cellular invasion in different microenvironments

Cadomian protolith ages of exotic mega blocks from Bugaj and Andrychów (Western outer Carpathians, Poland) and their palaeogeographic significance

This study presents the first zircon U-Pb LA-MC-ICP-MS ages and whole-rock Rb/Sr and Sm/Nd data from exotic blocks (Bugaj and Andrychów) from the Western Outer Carpathians (WOC) flysch. The CL images of the zircon crystals from both samples reveal typical magmatic textures characterized by a well-defined concentric and oscillatory growth zoning. A concordia age 580.1 ± 6.0 Ma of the zircons from the Bugaj sample is considered to represent the crystallization age of this granite. The zircon crystals from the Andrychów orthogneiss yield an age of 542 ± 21 Ma, in-terpreted as the uppermost Proterozoic, magmatic crystallization age of the granitoid protholith. The initial (at ca. 580 Ma) 87Sr/86Sr ratios of the Bugaj granitoids (0.72997 and 0.72874) are highly radio-genic, pointing to the assimilation of an older, possibly strongly Rb enriched source to the Bugaj melt. The Nd isotope systematics (εNd580 –1.4 and 0.4) also point to a significant contribution of such a dis-tinct mantle source. On the basis of the sequence of magmatic events obtained from U-Pb zircon ages, we suggest that exotic mega blocks deposited to the WOC basins were related to the Brunovistulicum Terrane. They belong to the group of Vendian/Cambrian granitoids representing the latest, post-tectonic expression of the Cadomian cycle

Geologiczne i geomorfologiczne uwarunkowania lokalizacji dawnych młynów wodnych nad dolną Liswartą

The location of the former water mills was studied on the lower Liswarta River on the Wieluń Upland. Fieldworks were supplemented with an analysis of the digital elevation model, detailed geological maps and historical topographic maps. The location of seven water mills was analysed with reference to local geological structures and limestone outcrops, as well as to the relief features of the Liswarta valley. Studies showed that water mills had been located usually in places, where the slope of the river channel increases. Such conditions correspond to mixed bedrock-alluvial segments of the river channel and narrow parts of the valley. There were no millponds on the lower reaches of the Liswarta River – only low weirs in the river channel were built. River discharge proved to be enough for working mills

Anthropogenic causes of wetland loss and degradation in the lower Kłodnica valley (southern Poland)

Loss and degradation of wetlands is now one of the most important environmental issues on a global scale. Previous research based on analyses of cartographic materials allow for quantification of changes in wetland area in recent centuries. The results of lithological research of peat cores, reported in this publication, have established that the processes of anthropogenic loss of wetlands can be much older and in the Kłodnica valley were initiated in the first millennium BC. As a result of increased mineral sedimentation accompanying soil erosion some peatlands have been fossilized whilst the area of others has been reduced. In total, the surface area of peat-forming wetlands in the bottom of the Kłodnica valley decreased by over 60% between the time of the Lusatian Culture settlement and the Middle Ages. Post-peatland habitats are recently used for agricultural or colonized by non-peat forming vegetation. These processes have played a more important role in the degradation of peatland ecosystems than the direct human impact in historic times. Changes in hydrographic networks, land drainage and regulation of water levels in rivers and canals in the last century have contributed to further reducing the wetland areas by almost 50% compared to the 1880s. These processes, however, have mainly affected ephemeral non-peat forming wetlands

Human-planted alder trees as a protection against debris flows (a dendrochronological study from the Moxi Basin, Southwestern China)

Large debris flows have destroyed the infrastructure and caused the death of people living in the Moxi Basin (Sichuan Province, Southwestern China). Inhabitants of the Moxi Basin live on the flat surfaces of debris-flow fans, which are also attractive for farming. During the monsoon season debris flows are being formed above the fans. Debris flows can destroy the houses of any people liv-ing within the fan surfaces. In order to prevent the adverse effects of flows, people plant alder trees (Alnus nepalensis) at the mouths of debris flow gullies running above debris flow fans. Alders are able to capture the debris transported during flow events. Trees are well adapted to surviving in con-ditions of environmental stress connected with abrupt transport and deposition of sediment from de-bris flows. Numerous wounds, tilting and bending of alder trees caused by debris flows only very rarely cause the death of trees. By dating scars and dating the time of alder tilting (through the analy-sis of annual rings), we have determined the frequency of debris flows occurring at the mouth of the Daozhao valley. In 1980-2012 within the studied debris-flow fan and the Daozhao gully, 2 large de-bris flow events occurred (1996, 2005) and some smaller events were probably recorded every 2-3 years

Basal localization of MT1-MMP is essential for epithelial cell morphogenesis in 3D collagen matrix

The membrane-anchored collagenase membrane type 1 matrix metalloprotease (MT1-MMP) has been shown to play an essential role during epithelial tubulogenesis in 3D collagen matrices; however, its regulation during tubulogenesis is not understood. Here, we report that degradation of collagen in polarized epithelial cells is post-translationally regulated by changing the localization of MT1-MMP from the apical to the basal surface. MT1-MMP predominantly localizes at the apical surface in inert polarized epithelial cells, whereas treatment with HGF induced basal localization of MT1-MMP followed by collagen degradation. The basal localization of MT1-MMP requires the ectodomains of the enzyme because deletion of the MT-loop region or the hemopexin domain inhibited basal localization of the enzyme. TGFβ is a well-known inhibitor of tubulogenesis and our data indicate that its mechanism of inhibition is, at least in part, due to inhibition of MT1-MMP localization to the basal surface. Interestingly, however, the effect of TGFβ was found to be bi-phasic: at high doses it effectively inhibited basal localization of MT1-MMP, whereas at lower doses tubulogenesis and basal localization of MT1-MMP was promoted. Taken together, these data indicate that basal localization of MT1-MMP is a key factor promoting the degradation of extracellular matrix by polarized epithelial cells, and that this is an essential part of epithelial morphogenesis in 3D collagen

Geneza i zasięg vistuliańskiego zespołu terasowego Odry-Osobłogi pod Krapkowicami

The system of morphological levels occurs on the slopes of the Odra and Osobłoga valleys in the vicinity of Krapkowice, southern Poland. Their genesis, range and age have been the subject of lively scientific discussion since the 1930s. Geomorphological analysis confirmed the fluvial origin of these flats. The use of LIDAR data allowed for precise height determination and revision of the range of indivi-dual river terraces, which rise above the valley bottom to a height of about 166 m a.s.l. (more or less 6 m above the Osobłoga/Odra channel), 168 m a.s.l. (8 m), 172 m a.s.l. (12 m), 174 m a.s.l. (14 m) and 181 m a.s.l. (21 m) res-pectively. The dating of sediments building the terrace elevated to 172 m a.s.l. using the OSL-SAR method gave a result of 87.7 ± 5.7 ka (GdTL-2820) indicating that it originated from the last cold stage of the Pleistocene. The ter-race surface was shaped by a sand-gravel braided river, which has developed on the substratum of the alluvium do-cumenting the period of intense valley-floor aggradation

Współczesne wykorzystanie przez bobra europejskiego Castor fiber antropogenicznie przekształconych dolin rzecznych (przykłady z Równiny Opolskiej i Wyżyny Woźnicko-Wieluńskiej)

W pracy przedstawiono typowe przypadki konstruktorskiej działalności bobrów w dolinach Małej Panwi i Liswarty oraz ich dopływów. Przeprowadzone badania wskazują, że bobry chętniej zasiedlają małe rzeki 3-4 rzędu niż rzeki główne. Najliczniejsze ślady działalności bobrów zaobserwowano na zalesionych odcinkach dolin Leńcy i Olszynki, gdzie gryzonie przekształciły 28-35% długości koryt rzecznych. Interesującym, niemal symbolicznym przypadkiem jest „naprawa” przez bobry grobli ziemnych starych stawów antropogenicznych i utworzenie w ich miejscu stawów bobrowych

Historical water-powered ferrous metallurgy reconstructed from tree-rings and lacustrine deposits (Mała Panew basin, southern Poland)

According to historical sources in the basin of Mała Panew River there were at least 56 water- powered iron smelters from 14th–19th century. Now only two metallurgy plants work in the area. Many of the former smelting settlements ceased to exist. Historical data on the smelting industry in the area are often scarce. The aim of the study was to reconstruct the history of ferrous metallurgy from (1) the remains of wooden historical buildings, (2) remains of charcoal kilns and (3) deposits from former smelter pond. Results show that Regolowiec smelting settlement existed already in the 17th century (at least several decades earlier than historical written sources suggest) and was later repaired after destruction caused probably by floods. Charcoal used for iron smelting in the ironworks in Brusiek on the Mała Panew River was burnt at the turn of the 18th century. This is in accordance with historical sources indicating particular prosperity of the metallurgy in that period. Upstream of the ironworks in Brusiek in the first half of the 17th century a large pond existed flooding the floor of the Mała Panew valley. Study has shown that the pond was at least 100 years older than historical sources have indicated

Characterization and regulation of MT1‐MMP cell surface‐associated activity

Quantitative assessment of MT1‐MMP cell surface‐associated proteolytic activity remains undefined. Presently, MT1‐MMP was stably expressed and a cell‐based FRET assay developed to quantify activity toward synthetic collagen‐model triple‐helices. To estimate the importance of cell surface localization and specific structural domains on MT1‐MMP proteolysis, activity measurements were performed using a series of membrane‐anchored MT1‐MMP mutants and compared directly with those of soluble MT1‐MMP. MT1‐MMP activity (kcat/KM) on the cell surface was 4.8‐fold lower compared with soluble MT1‐MMP, with the effect largely manifested in kcat. Deletion of the MT1‐MMP cytoplasmic tail enhanced cell surface activity, with both kcat and KM values affected, while deletion of the hemopexin‐like domain negatively impacted KM and increased kcat. Overall, cell surface localization of MT1‐MMP restricts substrate binding and protein‐coupled motions (based on changes in both kcat and KM) for catalysis. Comparison of soluble and cell surface‐bound MT2‐MMP revealed 12.9‐fold lower activity on the cell surface. The cell‐based assay was utilized for small molecule and triple‐helical transition state analog MMP inhibitors, which were found to function similarly in solution and at the cell surface. These studies provide the first quantitative assessments of MT1‐MMP activity and inhibition in the native cellular environment of the enzyme.MT1‐MMP was stably expressed and a cell‐based FRET assay developed to quantify activity toward synthetic collagen‐model triple‐helices. Activity measurements were performed using a series of membrane‐anchored MT1‐MMP mutants and compared directly with those of soluble MT1‐MMP. Cell surface localization of MT1‐MMP was found to restrict substrate binding and protein‐coupled motions for catalysis. Small molecule and triple‐helical transition state analog MMP inhibitors were found to function similarly in solution and at the cell surface.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/150520/1/cbdd13450.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/150520/2/cbdd13450_am.pd