Comparative structural analysis of a novel glutathioneS-transferase (ATU5508) from Agrobacterium tumefaciens at 2.0 A resolution

Glutathione S-transferases (GSTs) comprise a diverse superfamily of enzymes found in organisms from all kingdoms of life. GSTs are involved in diverse processes, notably small-molecule biosynthesis or detoxification, and are frequently also used in protein engineering studies or as biotechnology tools. Here, we report the high-resolution X-ray structure of Atu5508 from the pathogenic soil bacterium Agrobacterium tumefaciens (atGST1). Through use of comparative sequence and structural analysis of the GST superfamily, we identified local sequence and structural signatures, which allowed us to distinguish between different GST classes. This approach enables GST classification based on structure, without requiring additional biochemical or immunological data. Consequently, analysis of the atGST1 crystal structure suggests a new GST class, distinct from previously characterized GSTs, which would make it an attractive target for further biochemical studies.     

The prevalence of incidentaloma--asymptomatic thyroid nodules in the Tricity (Gdansk, Sopot, Gdynia) population

INTRODUCTION: The increased sensitivity of imaging devices raised number of incidentally discovered lesions in various organs of the human body. Thyroid gland is one of them. Reported prevalence of ultrasonographically detected thyroid nodules (incidentalomas) in general population ranges from 5.2 to 67.0%. Our study demonstrated occurrence of this clinical problem in the general population of the Tricity. MATERIAL AND METHODS: 135 healthy adults (95 women and 40 men) were examined. Neck palpation, ultrasonographic examinations of thyroid gland and serum tyreotropin (TSH) level measurement were made. RESULTS: In 8.9% of examine (12/135) persons nodules were palpable whereas in 14.8% (20/135) they were detectable only in ultrasonographic examination. Altogether thyroid ultrasound and palpation revealed nodules in 23.7% (32/135) of all cases. Multiple nodules were present in 12.0% of the cases. The pathology was more common in the elderly and in women. TSH serum level was within normal range in all cases of incidentaloma with otherwise normal thyroid gland. CONCLUSIONS: Prevalence of thyroid gland nodules (palpation--8.9% plus ultrasonography--14.8%) in healthy population of Gdansk, Gdynia and Sopot is close to data reported in southern Finland (27.0%) and Belgium (19.0%), where iodine deficiency is small, like in the Tricity area. The revealed lesions were over two times more frequent in the female population. Most of the nodules were not palpable.     

A novel mammalian HORMA domain-containing protein, HORMAD1, preferentially associates with unsynapsed meiotic chromosomes

HORMA domain-containing proteins regulate interactions between homologous chromosomes (homologs) during meiosis in a wide range of eukaryotes. We have identified a mouse HORMA domain-containing protein, HORMAD1, and biochemically and cytologically shown it to be associated with the meiotic chromosome axis. HORMAD1 first accumulates on the chromosomes during the leptotene to zygotene stages of meiotic prophase I. As germ cells progress into the pachytene stage, HORMAD1 disappears from the synapsed chromosomal regions. However, once the chromosomes desynapse during the diplotene stage, HORMAD1 again accumulates on the chromosome axis of the desynapsed homologs. HORMAD1 thus preferentially localizes to unsynapsed or desynapsed chromosomal regions during the prophase I stage of meiosis. Analysis of mutant strains lacking different components of the synaptonemal complex (SC) revealed that establishment of the SC is required for the displacement of HORMAD1 from the chromosome axis. Our results therefore strongly suggest that also mammalian cells use a HORMA domain-containing protein as part of a surveillance system that monitors synapsis or other interactions between homologs.     

Human mitochondrial RNA turnover caught in flagranti: involvement of hSuv3p helicase in RNA surveillance

The mechanism of human mitochondrial RNA turnover and surveillance is still a matter of debate. We have obtained a cellular model for studying the role of hSuv3p helicase in human mitochondria. Expression of a dominant-negative mutant of the hSUV3 gene which encodes a protein with no ATPase or helicase activity results in perturbations of mtRNA metabolism and enables to study the processing and degradation intermediates which otherwise are difficult to detect because of their short half-lives. The hSuv3p activity was found to be necessary in the regulation of stability of mature, properly formed mRNAs and for removal of the noncoding processing intermediates transcribed from both H and L-strands, including mirror RNAs which represent antisense RNAs transcribed from the opposite DNA strand. Lack of hSuv3p function also resulted in accumulation of aberrant RNA species, molecules with extended poly(A) tails and degradation intermediates truncated predominantly at their 3′-ends. Moreover, we present data indicating that hSuv3p co-purifies with PNPase; this may suggest participation of both proteins in mtRNA metabolism.     

MMP-12 Deficiency Attenuates Angiotensin II-Induced Vascular Injury,M2 Macrophage Accumulation,and Skin and Heart Fibrosis

MMP-12, a macrophage-secreted elastase, is elevated in fibrotic diseases, including systemic sclerosis (SSc) and correlates with vasculopathy and fibrosis. The goal of this study was to investigate the role of MMP-12 in cardiac and cutaneous fibrosis induced by angiotensin II infusion. Ang II-induced heart and skin fibrosis was accompanied by a marked increase of vascular injury markers, including vWF, Thrombospondin-1 (TSP-1) and MMP-12, as well as increased number of PDGFRβ⁺ cells. Furthermore Ang II infusion led to an accumulation of macrophages (Mac3⁺) in the skin and in the perivascular and interstitial fibrotic regions of the heart. However, alternatively activated (Arg 1⁺) macrophages were mainly present in the Ang II infused mice and were localized to the perivascular heart regions and to the skin, but were not detected in the interstitial heart regions. Elevated expression of MMP-12 was primarily found in macrophages and endothelial cells (CD31⁺) cells, but MMP-12 was not expressed in the collagen producing cells. MMP-12 deficient mice (MMP12KO) showed markedly reduced expression of vWF, TSP1, and PDGFRβ around vessels and attenuation of dermal fibrosis, as well as the perivascular fibrosis in the heart. However, MMP-12 deficiency did not affect interstitial heart fibrosis, suggesting a heterogeneous nature of the fibrotic response in the heart. Furthermore, MMP-12 deficiency almost completely prevented accumulation of Arg 1⁺ cells, whereas the number of Mac3⁺ cells was partially reduced. Moreover production of profibrotic mediators such as PDGFBB, TGFβ1 and pSMAD2 in the skin and perivascular regions of the heart was also inhibited. Together, the results of this study show a close correlation between vascular injury markers, Arg 1⁺ macrophage accumulation and fibrosis and suggest an important role of MMP-12 in regulating these processes.     

Human platelets release alpha-6-L-fucosyltransferase upon activation

Previously we have shown that human platelets release alpha-6-L-fucosyltransferase (EC 2.4.1.68) during coagulation of blood [(1987) Glycoconjugate J. 4, 43-49]. Here we report that agonists which induce platelet aggregation bring about release of the enzyme. In quantitative terms the release of alpha-6-L-fucosyltransferase by washed, aggregated platelets was very similar to that occurring during blood coagulation.