Dimeric novel HSP40 is incorporated into the radial spoke complex during the assembly process in flagella

The radial spoke is a stable structural complex in the 9 + 2 axoneme for the control of flagellar motility. However, the spokes in Chlamydomonas mutant pf24 are heterogeneous and unstable, whereas several spoke proteins are reduced differentially. To elucidate the defective mechanism, we clone RSP16, a prominent spoke protein diminished in pf24 axonemes. Unexpectedly, RSP16 is a novel HSP40 member of the DnaJ superfamily that assists chaperones in various protein-folding-related processes. Importantly, RSP16 is uniquely excluded from the 12S spoke precursor complex that is packaged in the cell body and transported toward the flagellar tip to be converted into mature 20S axonemal spokes. Rather, RSP16, transported separately, joins the precursor complex in flagella. Furthermore, RSP16 molecules in vitro and in flagella form homodimers, a characteristic required for the cochaperone activity of HSP40. We postulate that the spoke HSP40 operates as a cochaperone to assist chaperone machinery at the flagellar tip to actively convert the smaller spoke precursor and itself into the mature stable complex; failure of the interaction between the spoke HSP40 and its target polypeptide results in heterogeneous unstable radial spokes in pf24.     

Shifted from Wnt to Hedgehog signaling pathways

Two papers in recent issue of Developmental Cell (Glise et al. 2005; Gorfinkiel et al. 2005) have shown that Shifted, a Drosophila ortholog of Wnt Inhibitory Factor (WIF), modulates the distribution of Hedgehog protein in the wing imaginal disc through a Wnt-independent mechanism.     

Negative regulation of monocyte adhesion to arterial elastic laminae by signal regulatory protein alpha and Src homology 2 domain-containing protein-tyrosine phosphatase-1

Elastic laminae are extracellular matrix constituents that not only contribute to the stability and elasticity of arteries but also play a role in regulating arterial morphogenesis and pathogenesis. We demonstrate here that an important function of arterial elastic laminae is to prevent monocyte adhesion, which is mediated by the inhibitory receptor signal regulatory protein (SIRP) alpha and Src homology 2 domain-containing protein-tyrosine phosphatase (SHP)-1. In a matrix-based arterial reconstruction model in vivo, elastic laminae were resistant to leukocyte adhesion and transmigration compared with the collagen-dominant arterial adventitia. The density of leukocytes within the elastic lamina-dominant media was about 58-70-fold lower than that within the adventitia from 1 to 30 days. An in vitro assay confirmed the inhibitory effect of elastic laminae on monocyte adhesion. The exposure of monocytes to elastic laminae induced activation of SIRP alpha, which in turn activated SHP-1. Elastic lamina degradation peptides extracted from arterial specimens could also activate SIRP alpha and SHP-1. The knockdown of SIRP alpha and SHP-1 by specific small interfering RNA diminished the inhibitory effect of elastic laminae, resulting in a significant increase in monocyte adhesion. These observations suggest that SIRP alpha and SHP-1 potentially mediate the inhibitory effect of elastic laminae on monocyte adhesion.     

Vialinin A, a novel 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenger from an edible mushroom in China

While screening for bioactive compounds from edible mushrooms, a new potent antioxidant, vialinin A (1), together with a known compound, ganbajunin B (2), and a mixture of ganbajunins D (3) and E (4), were isolated from the dry fruiting bodies of Thelephora vialis. The structure of 1, 5',6'-bis(phenylacetoxy)-1,1':4',1'-terphenyl-2',3',4,4'-tetraol, was elucidated by spectroscopic and chemical methods. This compound had strong 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical-scavenging activity with an EC(50) value of 14.0 microM, nearly equal to that of butylated hydroxytoluene (BHT; EC(50) = 10.0 microM). A radical scavenging experiment using 1 and DPPH radicals indicated that 1 donated two hydrogen atoms to two molecules of the DPPH radical under hydrophobic conditions.     

Expression and secretion of the protective antigen of Bacillus anthracis in Bacillus brevis

We used the Bacillus brevis-pNU212 system to develop a mass production system for the protective antigen (PA) of Bacillus anthracis. A moderately efficient expression-secretion system for PA was constructed by fusing the PA gene from B. anthracis with the B. brevis cell-wall protein signal-peptide encoding region of pNU212, and by introducing the recombinant plasmid, pNU212-mPA, into B. brevis 47-5Q. The clone producing PA secreted about 300 microg of recombinant PA (rPA) per ml of 5PY-erythromycin medium after 4 days incubation at 30 degrees C. The rPA was fractionated from the culture supernatant of B. brevis 47-5Q carrying pNU212-mPA using ammonium sulfate at 70% saturation followed by anion exchange chromatography on a Hitrap Q, a Hiload 16/60 Superdex 200 gel filtration column and a phenyl sepharose hydrophobic interaction column, yielding 70 mg rPA per liter of culture. The N-terminal sequence of the purified rPA was identical to that of native PA from B. anthracis. The purified rPA exhibited cytotoxicity towards J774A.1 cells when combined with lethal factor. The rPA formulated in either Rehydragel HPA or MPL-TDM-CWS adjuvant (Ribi-Trimix) elicited the expression of a large amount of anti-PA and neutralizing antibodies in guinea pigs and completely protected them against a 100 LD50 challenge with fully virulent B. anthracis spores.     

Myosin filament assembly in an ever-changing myofilament lattice of smooth muscle

A major development in smooth muscle research in recent years is the recognition that the myofilament lattice of the muscle is malleable. The malleability appears to stem from plastic rearrangement of contractile and cytoskeletal filaments in response to stress and strain exerted on the muscle cell, and it allows the muscle to adapt to a wide range of cell lengths and maintain optimal contractility. Although much is still poorly understood, we have begun to comprehend some of the basic mechanisms underlying the assembly and disassembly of contractile and cytoskeletal filaments in smooth muscle during the process of adaptation to large changes in cell geometry. One factor that likely facilitates the plastic length adaptation is the ability of myosin filaments to form and dissolve at the right place and the right time within the myofilament lattice. It is proposed herein that formation of myosin filaments in vivo is aided by the various filament-stabilizing proteins, such as caldesmon, and that the thick filament length is determined by the dimension of the actin filament lattice. It is still an open question as to how the dimension of the dynamic filament lattice is regulated. In light of the new perspective of malleable myofilament lattice in smooth muscle, the roles of many smooth muscle proteins could be assigned or reassigned in the context of plastic reorganization of the contractile apparatus and cytoskeleton.     

Human and murine paraoxonase 1 are host modulators of Pseudomonas aeruginosa quorum-sensing

The pathogenic bacterium Pseudomonas aeruginosa uses acyl-HSL quorum-sensing signals to regulate genes controlling virulence and biofilm formation. We found that paraoxonase 1 (PON1), a mammalian lactonase with an unknown natural substrate, hydrolyzed the P. aeruginosa acyl-HSL 3OC12-HSL. In in vitro assays, mouse serum-PON1 was required and sufficient to degrade 3OC12-HSL. Furthermore, PON2 and PON3 also degraded 3OC12-HSL effectively. Serum-PON1 prevented P. aeruginosa quorum-sensing and biofilm formation in vitro by inactivating the quorum-sensing signal. Although 3OC12-HSL production by P. aeruginosa was important for virulence in a mouse sepsis model, Pon1-knock-out mice were paradoxically protected. These mice showed increased levels of PON2 and PON3 mRNA in epithelial tissues suggesting a possible compensatory mechanism. Thus, paraoxonase interruption of bacterial communication represents a novel mechanism to modulate quorum-sensing by bacteria. The consequences for host immunity are yet to be determined.     

记贵州关岭生物群中的大型鱼龙Shastasaurus(英文)

贵州关岭三叠纪法郎组瓦窑段除产出大量保存完整的海百合化石外,还产出多门类海生爬行动物化石。迄今为止,关岭生物群已报道的中-大型鱼龙类有6属6种,包括邓氏贵州鱼龙(Guizhouichthyosaurus tangae)、蔡胡氏典型鱼龙(Typicusichthyosaurus tsaihuae)、梁氏关岭鱼龙(Guanlingsaurus liangae)、亚洲杯椎鱼龙(Cymbospondylus asiaticus)、美丽盘江龙(Panjiangsaurus epicharis)和卧龙岗卡洛维龙(Callawayia wolonggangense)。一些种属的存在长期以来争议较大,目前多数观点倾向将Cymbospondylus asiaticus和Panjiangsaurus epicharis归并于Guizhouichthyosaurus tangae,将Typicusichthyosaurus tsaihuae归并于Guanlingsaurus liangae。Guanlingsaurus以具有较短的吻部和较多的荐前椎数目而与Guizhouichthyosaurus有明显的不同(Maisch et al.,2006)。虽然Maisch et al.(2006)对G.tangae进行了重新研究,但他们的研究重点是头部骨骼,有限的头后骨骼信息来自一具未经充分修理的骨架(GNP-d41),一些特征未能清晰揭示。头后骨骼材料的缺乏限制了对该种分类位置的判断(Maisch et al., 2006)。在中、晚三叠世大型鱼龙中,肩带和腰带骨骼的形态、前肢和后肢骨骼的特征常具有非常重要的系统分类学意义。本文通过对关岭生物群的一具保存完整的大型鱼龙骨骼化石的详细研究,对Guizhouichthyosaurus tangae的归属进行了重新修订。研究标本(IVPP V 11853)产于贵州省关岭县新铺乡法郎组瓦窑段。修理后的骨架全长5.2 m,以腹面向上保存,尾部后部以右侧面向上保存。除前、后肢部分残缺外,其他部位均保存完整,肩带骨骼和腰带骨骼均原位保存。头骨背腹向压扁,仅右侧角的上颞骨有部分破损。新材料的头后骨骼特征表明该种应该属于萨斯特鱼龙科(Shastasauridae)的萨斯特鱼龙属(Shastasaurus)。Shastasaurus是一个广为人知的晚三叠世大型鱼龙化石属,最初建立于美国加利福尼亚州的Shasta地区,包括5个种(Merriam,1895,1902,1908)。由于建立这些种的化石材料均保存不好,尤其是模式种仅建立于几块背椎、背肋和两块耻骨之上,导致有关萨斯特鱼龙属和萨斯特鱼龙科的一些特征定义不明确。后人倾向于将已建立的种进行归并,即将Shastasaurus alexandrae、S.altispinus和S.osmonti划归模式种S.pacificus,将S.careyi另归入Shonisaurus sp.。虽然萨斯特鱼龙类的其他属,如Shonisaurus、Besanosaurus、Metashastasaurus、Pessosaurus等,在加拿大、墨西哥、意大利、瑞士等地的中、晚三叠世地层中被陆续发现和报道,但除美国加利福尼亚州的材料外,其他地区发现的萨斯特鱼龙属的化石均未得到广泛认可。本文将G.tangae归入萨斯特鱼龙属主要依据对该种模式标本(Gmr 009)的重新观察和对该种一新材料(IVPP V 11853)头后骨骼的深入研究。两骨架的头后部分尤其是前肢和肩带均表现出与已知的萨斯特鱼龙属各种的极大相似性,具体包括:椎体稍短;颈肋近端双头;尾椎腹侧Y型人字骨发育;锁骨细长,中部向后弯;乌喙骨斧状,中部收缩形成前后缘不对称的"茎";肩胛骨呈宽的镰刀状,前缘强烈外展;肱骨、桡骨、尺骨均较短;肱骨和桡骨前缘具凹缺;桡骨明显大于尺骨,桡骨后缘和尺骨前缘略凹入;相对较大的桡腕骨等。腰带和后肢特征与萨斯特鱼龙类其他属差别不大。除头后骨骼特征外,关岭材料的头骨特征也与萨斯特鱼龙属惟一的一个保存有部分头骨骨骼的S.alexandrae的特征大体相符,如具有大的、前后略拉长的眼眶,相对较窄的颊部,横向伸展的鼻骨和额骨的接触面,颞孔稍小于眼眶等。同时,通过与萨斯特鱼龙类其他各属化石的对比研究发现,虽然一些研究者认为萨斯特鱼龙属的荐前椎数目在55个左右,且指(趾)骨为3指(趾),但鉴于已经发现同属于萨斯特鱼龙类的Shonisaurus、Besanosaurus、Metashastasaurus等属或者具有超过60的荐前椎数,或者指(趾)骨为4指(趾),同时迄今为止尚无完整的萨斯特鱼龙属的化石标本被发现,有关其荐前椎数目和指(趾)骨列数仅是推测,因此基于中国贵州关岭的标本,本文认为萨斯特鱼龙属很可能具有超过60的荐前椎数,同时指骨(趾骨)是4指(趾)。在观察和对比了众多产于关岭的大型鱼龙骨骼化石的基础上,将邓氏贵州鱼龙、亚洲杯椎鱼龙和美丽盘江龙重新厘定为邓氏萨斯特鱼龙,并对该种的头后骨骼特征进行了详细描述。这是萨斯特鱼龙属的分子第一次在中国被确认,为了解该属鉴定特征和古地理分布提供了新的信息。