Tat-specific cytotoxic T lymphocytes select for SIV escape variants during resolution of primary viraemia

Human immunodeficiency virus (HIV) and simian immunodeficiency virus (SIV) infections are characterized by early peaks of viraemia that decline as strong cellular immune responses develop. Although it has been shown that virus-specific CD8-positive cytotoxic T lymphocytes (CTLs) exert selective pressure during HIV and SIV infection, the data have been controversial. Here we show that Tat-specific CD8-positive T-lymphocyte responses select for new viral escape variants during the acute phase of infection. We sequenced the entire virus immediately after the acute phase, and found that amino-acid replacements accumulated primarily in Tat CTL epitopes. This implies that Tat-specific CTLs may be significantly involved in controlling wild-type virus replication, and suggests that responses against viral proteins that are expressed early during the viral life cycle might be attractive targets for HIV vaccine development.     

Fringe is a glycosyltransferase that modifies Notch

Notch receptors function in highly conserved intercellular signalling pathways that direct cell-fate decisions, proliferation and apoptosis in metazoans. Fringe proteins can positively and negatively modulate the ability of Notch ligands to activate the Notch receptor. Here we establish the biochemical mechanism of Fringe action. Drosophila and mammalian Fringe proteins possess a fucose-specific beta1,3 N-acetylglucosaminyltransferase activity that initiates elongation of O-linked fucose residues attached to epidermal growth factor-like sequence repeats of Notch. We obtained biological evidence that Fringe-dependent elongation of O-linked fucose on Notch modulates Notch signalling by using co-culture assays in mammalian cells and by expression of an enzymatically inactive Fringe mutant in Drosophila. The post-translational modification of Notch by Fringe represents a striking example of modulation of a signalling event by differential receptor glycosylation and identifies a mechanism that is likely to be relevant to other signalling pathways.     

鱼类饲料配方及制作

在约300种养殖鱼类中,仅50种有营养学数据,不到10 种有完全的营养学数据.在饲料蛋白源中,鱼粉是鲑鳟鱼类饲料的主要蛋白源,蛋白质含量为60%～80 %.肉骨粉一般蛋白质含量为50 %左右,赖氨酸含量较鱼粉低.肉骨粉灰分含量可到30 %,限制了其在鱼类饲料中的应用.一般在鲑鳟鱼类饲料中用量不超过10 %,温水性鱼类饲料中用量不超过15 %.血粉蛋白质含量为80 %～86 %,是很好的赖氨酸来源,但缺乏蛋氨酸.血粉的适口性较差,在鲑鳟饲料中用量不超过10 %,温水性鱼类饲料中不超过5 %.肉骨-血粉混合物的营养成分与鱼粉接近,可完全替代沟鲶饲料中的鱼粉.鸡肉粉蛋白质含量为60 %～70 %,可替代鲑鳟饲料中10 %的鱼粉及温水性鱼类饲料中50 %的蛋白质.水解羽毛粉蛋白质含量约为85 %,但消化率较低.可替代鲑鳟及温水性鱼类饲料中10 %的蛋白质.鸡肉粉-羽毛粉混合物可替代虹鳟饲料中50 %的鱼粉.豆粕可替代虹鳟饲料中50 %的鱼粉蛋白,是温水性鱼类饲料的主要蛋白源.豆粕中存在一些抗营养因子.一般豆油提取过程可以分解大部分抗营养因子.一些报道表明进一步热处理可以提高豆粕的营养价值.经过热处理的全脂豆粉,如果添加蛋氨酸及赖氨酸,可以替代虹鳟及鲤鱼饲料中50 %～75 %的鱼粉.棉粕对鱼的适口性好,但缺少赖氨酸.除非添加赖氨酸,棉粕在温水性鱼类饲料中用量不应超过30 %.花生粕适口性好,但缺少赖氨酸.添加量超过20 %时,需添加赖氨酸.葵花子粕赖氨酸含量较低,可替代虹鳟饲料中40%的鱼粉及温水性鱼类饲料中20 %的鱼粉.低毒菜粕可替代虹鳟饲料中66 %的鱼粉.添加增味剂后,去植酸菜籽蛋白浓缩物可完全替代虹鳟饲料中的鱼粉.玉米面筋含42 %～60 %的蛋白质,添加水平在虹鳟饲料可达20 %,鲤鱼饲料可达10 %.酒精业副产物含27 %蛋白质,适口性好但赖氨酸含量低,添加水平在虹鳟饲料可达10 %,温水性鱼类饲料可达30 %.非营养因子也会影响到饲料制作.如制作膨化饲料需至少25 %的谷类原料.棉粕含量过高会减缓挤压速度.非营养性添加剂包括黏合剂、类胡萝卜素、鱼药、激素、抗微生物及真菌制剂、抗氧化剂、增味剂,但不应在饲料中添加纤维素,其含量应尽量降低.饲料制作工艺包括蒸汽制粒及挤压.挤压可制作浮性、沉性及缓沉性饲料,并能提高消化率,但会分解一些维生素.     

Hypervariable 'minisatellite' regions in human DNA

The human genome contains many dispersed tandem-repetitive 'minisatellite' regions detected via a shared 10-15-base pair 'core' sequence similar to the generalized recombination signal (chi) of Escherichia coli. Many minisatellites are highly polymorphic due to allelic variation in repeat copy number in the minisatellite. A probe based on a tandem-repeat of the core sequence can detect many highly variable loci simultaneously and can provide an individual-specific DNA 'fingerprint' of general use in human genetic analysis.     

Substrate specificity and affinity of a protein modulated by bound water molecules

Water molecules influence molecular interactions in all biological systems, yet it is extremely difficult to understand their effects in precise atomic detail. Here we present evidence, based on highly refined atomic structures of the complexes of the L-arabinose-binding protein with L-arabinose, D-fucose and D-galactose, that bound water molecules, coupled with localized conformational changes, can govern substrate specificity and affinity. The atoms common to the three sugars are identically positioned in the binding site and the same nine strong hydrogen bonds are formed in all three complexes. Two hydrogen-bonded water molecules in the site contribute further to tight binding of L-arabinose but create an unfavourable interaction with the methyl group of D-fucose. Equally tight binding of D-galactose is attained by the replacement of one of the hydrogen-bonded water molecules by its--CH2OH group, coordinated with localized structural changes which include a shift and redirection of the hydrogen-bonding interactions of the other water molecule. These observations illustrate how ordered water molecules can contribute directly to the properties of proteins by influencing their interaction with ligands.     

A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast

A protein sensitive to N-ethylmaleimide catalyses the fusion of transport vesicles with Golgi cisternae in a mammalian cell-free system. By cloning and sequencing its gene from Chinese hamster ovary cells and by use of in vitro assays, we show that this fusion protein is equivalent to the SEC18 gene product of the yeast Saccharomyces cerevisiae, known to be essential for vesicle-mediated transport from the endoplasmic reticulum to the Golgi apparatus. The mechanism of vesicular fusion is thus highly conserved, both between species and at different stages of transport.