Half-att site substrates reveal the homology independence and minimal protein requirements for productive synapsis in lambda excisive recombination

The early events in site-specific excisive recombination were studied with phage lambda half-att sites that have no DNA to one side of the strand exchange region; they carry a single core-type integrase binding site and either P or P' arm flanking DNA. These half-attR and half-attL sites exhibit normal properties for the initial (covalent) top-strand transfer and form stable intermediates independent of later steps in the reaction. With these novel substrates we show that Xis specifically promotes the first strand exchange and that attL enhances Int cleavage at the top-strand site of attR. It is also shown that synapsis and initial strand transfers do not require DNA-DNA pairing but are mediated by protein-protein and protein-DNA interactions. These involve the two top-strand Int binding sites (required for the first strand exchange) and, in addition, one of the two bottom-strand sites (C') responsible for the second strand exchange.     

Evidence for a new biologic pathway of androstenedione synthesis from 11-deoxycortisol

17-Hydroxyprogesterone is a well-known precursor of androstenedione in adrenal biosynthesis. This study using sheep adrenal incubations demonstrates that 11-deoxycortisol, the precursor of cortisol synthesis, also can be a precursor of androstenedione. Indeed, our data show that androstenedione synthesis is negatively correlated to the synthesis of cortisol and cortisone. This fact allowed us to infer that this new pathway is closely related to the activity of the 11 beta-hydroxylase that is responsible for the synthesis of cortisol. Indeed, when the activity of this enzyme is impaired, 11-deoxycortisol follows the pathway that leads to androstenedione synthesis in the adrenals. This pathway could explain, at least in part, the marked increase of androstenedione observed in congenital adrenal hyperplasia due to 11 beta-hydroxylase deficiency.     

烟草花粉管亚原生质体的分离和培养行为

应用酶法从烟草花粉管分离出大量亚原生质体。具核的和无核的亚原生质体之比约为1:1。这种亚原生质体在D_2培养基中培养后,不论有核的或是无核的都能生长管状结构和再生厚的壁。管状结构的生长有节律性,常呈结节状。随着管状结构的生长,细胞内含物逐渐流入生长中的管状结构内,有时会从薄的管状结构的顶端排出到培养液中。已生长管状结构的亚原生质体,具核的和无核的比例约为1:1.7,表明管状结构的生长和壁的再生与是否有核的存在无关。对酶液处理后花粉管亚原生质体从花粉管的释放和从单独的花粉管亚原生质体生长管状结构的过程,进行了活体连续观察和照相记录。实验结果说明,结节状的管状结构确实是从单独的一个亚原生质体形成的。管状结构的生长和壁的再生似乎与细胞质进入新生的管状结构有关。讨论了花粉管亚原生质体在植物遗传操作中应用的可能性。     

Studies on Microbial Ribonucleic Acid VI. Appearance of Methyl-deficient Transfer Ribonucleic Acid During Logarithmic Growth of Saccharomyces cerevisiae

Transfer ribonucleic acid (tRNA) that is deficient in methyl groups may be detected in logarithmically growing Saccharomyces cerevisiae. The amount of methyl-deficient tRNA is not constant throughout the logarithmic phase, but is maximal about one generation before the onset of the late growth phase. During this latter phase, the tRNA is fully methylated. The methyl-deficient tRNA is present during a period of high metabolic activity of the cell, characterized by increased RNA and protein content.