Cellobiose-oxidizing enzyme from a newly isolated cellulolytic bacterium Cytophaga sp. LX-7

Summary The cellobiose oxidizing enzyme of the newly isolated cellulolytic bacterium Cytophaga sp. LX-7 was produced extracellularly when grown on cellulose or other saccharides, which was previously noted only in fungi. The enzyme could use not only cellobiose, maltose, glucose and other saccharides but also cellulose as substrates, and use dichlorophenol indophenol and oxygen as electron acceptors.     

Calcium binding to the subunit c ofE. coli ATP-synthase and possible functional implications in energy coupling

The 8-kDa subunit c of theE. coli F₀ ATP-synthase proton channel was tested for Ca⁺⁺ binding activity using a⁴⁵Ca⁺⁺ ligand blot assay after transferring the protein from SDS-PAGE gels onto polyvinyl difluoride membranes. The purified subunit c binds⁴⁵Ca⁺⁺ strongly with Ca⁺⁺ binding properties very similar to those of the 8-kDa CF₀ subunit III of choloroplast thylakoid membranes. The N-terminal f-Met carbonyl group seems necessary for Ca⁺⁺ binding capacity, shown by loss of Ca⁺⁺ binding following removal of the formyl group by mild acid treatment. The dicyclohexylcarbodiimide-reactive Asp-61 is not involved in the Ca⁺⁺ binding, shown by Ca⁺⁺ binding being retained in twoE. coli mutants, Asp61Asn and Asp61Gly. The Ca⁺⁺ binding is pH dependent in both theE. coli and thylakoid 8-kDa proteins, being absent at pH 5.0 and rising to a maximum near pH 9.0. A treatment predicted to increase the Ca⁺⁺ binding affinity to its F₀ binding site (chlorpromazine photoaffinity attachment) caused an inhibition of ATP formation driven by a base-to-acid pH jump in whole cells. Inhibition was not observed when the Ca⁺⁺ chelator EGTA was present with the cells during the chlorpromazine photoaffinity treatment. An apparent Ca⁺⁺ binding constant on the site responsible for the UV plus chlorpromazine effect of near 80–100 nM was obtained using an EGTA-Ca⁺⁺ buffer system to control free Ca⁺⁺ concentration during the UV plus chlorpromazine treatment. The data are consistent with the notion that Ca⁺⁺ bound to the periplasimic side of theE. coli F₀ proton channel can block H⁺ entry into the channel. A similar effect occurs in thylakoid membranes, but the Ca⁺⁺ binding site is on the lumen side of the thylakoid, where Ca⁺⁺ binding can modulate acid-base jump ATP formation. The Ca⁺⁺ binding to the F₀ and CF₀ complexes is consistent with a pH-dependent gating mechanism for control of H⁺ ion flux across the opening of the H⁺ channel.This work was supported in part by grants from the Department of Energy and the U.S. Department of Agriculture.On leave from the Institute of Soil Science and Photosynthesis, Russian Academy of Science, Pushchino, Russia.     

蓝氏贾第虫核被膜缺口的电镜观寨

双滴虫类是迄今所知的现存最原始的真核生物类群。以蓝氏贾第虫作为双滴虫类的代表,对其细胞核进行了电镜观察。除了未见有核仁外,还发现其核被膜的横切面上存在有缺口。在缺口的边缘处,核内膜与校外膜是相互连接着的,表明并非切片时所造成的假象。核被膜缺口处常有一核纤层样的薄层分隔核质与细胞质。用高锰酸钾固定细胞以求只保存膜结构时,核被膜缺口仍然可见,上述的薄层即未见到。核被膜缺口的发现证实了李靖炎（１９７９）的核被膜起源假说所作出的推断。     

The scl gene product is required for the generation of all hematopoietic lineages in the adult mouse.

Homozygosity for a null mutation in the scl gene causes mid-gestational embryonic lethality in the mouse due to failure of development of primitive hematopoiesis. Whilst this observation established the role of the scl gene product in primitive hematopoiesis, the death of the scl null embryos precluded analysis of the role of scl in later hematopoietic development. To address this question, we created embryonic stem cell lines with a homozygous null mutation of the scl gene (scl-/-) and used these lines to derive chimeric mice. Analysis of the chimeric mice demonstrates that the scl-/- embryonic stem cells make a substantial contribution to all non-hematopoietic tissues but do not contribute to any hematopoietic lineage. These observations reveal a crucial role for the scl gene product in definitive hematopoiesis. In addition, in vitro differentiation assays with scl-/- embryonic stem cells showed that the scl gene product was also required for formation of hematopoietic cells in this system.     

A splicing-dependent regulatory mechanism that detects translation signals.

Premature termination codons (PTCs) can cause the decay of mRNAs in the nuclear fraction of mammalian cells. This enigmatic nuclear response is of interest because it suggests that translation signals do not restrict their effect to the cytoplasm, where fully assembled ribosomes reside. Here we examined the molecular mechanism for this putative nuclear response by using the T-cell receptor-beta (TCR-beta) gene, which acquires PTCs as a result of programmed rearrangements that occur during normal thymic ontogeny. We found that PTCs had little or no measurable effect on TCR-beta pre-mRNA levels, but they sharply depressed TCR-beta mature mRNA levels in the nuclear fraction of stably transfected cells. A PTC split by an intron was able to trigger the down-regulatory response, implying that PTC recognition occurs after an mRNA is at least partially spliced. However, intron deletion and addition studies demonstrated that a PTC must be followed by at least one functional (spliceable) intron to depress mRNA levels. One explanation for this downstream intron-dependence is that cytoplasmic ribosomes adjacent to nuclear pores scan mRNAs still undergoing splicing as they emerge from the nucleus. We found this explanation to be unlikely because PTCs only 8 or 10 nt upstream of a terminal intron down-regulated mRNA levels, even though this distance is too short to permit PTC recognition in the cytoplasm prior to the splicing of the downstream intron in the nucleus. Collectively, the results suggest that nonsense codon recognition may occur in the nucleus.     

Cornucoryphe schirmi n. gen. n. sp., an unusual conocoryphid trilobite from the Middle Cambrian of Spain

Cornucoryphe n. gen. from the Middle Cambrian of the Cantabrian Mountains and Aragón is a unicornianConocoryphe with pronounced alae.     

Immunolocalization of extensin and potato tuber lectin in carrot, tomato and potato

Tissue-specific expression of two members of the cell wall hydroxyproline-rich glycoprotein (HRGP) family, extensin and potato tuber lectin, was examined by immunolocalization at the light microscope level in various organs (leaves, stems, roots, fruit, tuber) of carrot ( Daucus carota cv. Thumbelina), tomato ( Lycopersicon esclentum cv. Pixie Hybrid II), and potato ( Solanum tuberosum cv. Kennebec). Extensin was prominently expressed in vascular tissue, particularly xylem and also phloem, although virtually all cells displayed some degree of staining which varied as a function of the tissue, organ, and plant under study. Antibodies against potato tuber lectin (PTL) displayed a localization pattern similar to that observed for extensin; notably PTL did not stain cambium but did stain epithelial cells lining secretory cavities. These distribution patterns are consistent with a role for extensin, and possibly PTL, in providing mechanical support in tissues subjected to compression or torsional stress imparted by vascular growth, or by similar stress brought about by transport of vascular fluids.     

On the existence of stable pairing distributions

We formulate and analyze pair-formation models for multiple groups with general pairing rates and arbitrary mixing probabilities. Under the assumption of constant recruitment rates and equal average duration of all types of partnerships, we have shown that the dynamics are relatively simple because of the monotonicity properties of the dynamical system associated with the pairing/mixing of heterogeneous populations of male and female individuals. In fact, we have shown that the corresponding asymptotic stable paired distribution is given precisely by the asymptotic values of the matrices that prescribe the mixing/contact structure. In other words, if the sizes of the mixing subpopulations of males and females are asymptotically constant and if the average durations of partnerships are about the same regardless of type, then the matrices that describe the mixing between subpopulations also characterize the distribution of paired types. Alternatively, if the distribution of the average duration of relationships between individuals has a large variance then it may be impossible to detect any relationship between the mixing/contact structure and the observed distribution of paired types. The study of models with constant per-capita recruitment rates give rise to homogeneous systems of degree one. The analysis of the dynamics of pairs for models with exponentially growing populations of singles is complicated. So far, we are only able to classify the stability of all non-strictly positive boundary exponential solutions. From our incomplete analysis, it is not possible to detect necessary and sufficient conditions for the existence and stability of strictly interior exponential solutions. We cannot rule out the possibility of oscillations. The mathematical problems associated with the stability of exponential solutions of dynamical systems of degree one are of relevance in demography, epidemiology, and population dynamics.On leave from University of Alabama in Huntsville     

自养黄杆菌合成羟基丁酸和羟基戊酸共聚体的发酵研究

采用本实验室从土壤中分离到的一株自养黄杆菌进行了羟基丁酸和羟基戊酸共聚体〔P(HB-co-HV)〕的发酵试验。实验结果表明,该菌株是自养黄杆菌葡萄糖运输突变株,可以葡萄糖、果糖、蔗糖、麦芽糖、乙酸盐、乳酸盐和苹果酸盐作为唯一碳源,尤以葡萄糖和果糖效果最佳。硫酸铵、氯化铵和蛋白胨等不同氮源不影响其生长,却影响细胞中P(HB-co-HV)的含量和P(HB-co-HV)中HV/HB的比例。应用两阶段控制方式,经42h的补料分批发酵,细胞浓度达34.9g·L~(-1),P(HB-co-HV)浓度达25.28g·L~(-1)。细胞和P(HB-co-HV)生产速率系数分别为0.83g·L~(-1)”·h~(-1)和0.61g·L~(-1)·h~(-1)。以基质为基准的细胞得率系数(Yx/s)、产物得率系数(Yp/s)和以干细胞为基准的产物得率系数(Yp/x)分别为0.283(g/g)、0.174(g/g)和0.73(g/g)。改变培养基中碳氮源组分可将P(HB-co-HV)中HB的含量调节在24％～78％之间。     

15N resonance assignments of oxidized and reducedChromatium vinosum high-potential iron protein

The¹⁵N resonances in reduced and oxidizedChromatium vinosum high-potential iron protein have been assigned by use of¹H-¹H COSY spectra and¹H-¹⁵N HMQC, HMQC-COSY, and HMQC-NOESY spectra. Unambiguous assignment of 70 of 85 backbone¹⁵N resonances in the reduced protein and 62 of 85 resonances in the oxidized protein are made, as are 12 of 21 side-chain¹⁵N resonances.