孤儿受体TR2 mRNA在大鼠和恒河猴隐睾内生精细胞凋亡过程中的表达

孤儿受体TR2 mRNA在大鼠睾丸内特异定位于生精细胞,主要在减数分裂期的精母细胞和减数分裂后的圆形和长型精子细胞表达。在不同曲细精管间,TR2 mRNA表达有明显差异。大鼠隐睾手术后d3和d5,生精细胞出现凋亡,TR2 mRNA表达略有下降;到d7.5时,大量生精细胞发生凋亡,TR2 mRNA表达明显降低。从手术后d10起,检测不到TR2 mRNA的表达。手术后d15和d20的恒河猴隐睾中,仅有少数初级精母细胞表达TR2 mRNA,d30、45和60恒河猴隐睾基本不表达TR2 mRNA。

真核生物mRNA二级结构与内含子剪接

对68个外显子-内含子-外显子序列片段以及相应的外显子-外显子序列片段的二级结构进行分析后发现,内含子5^端和3^端的碱基G(剪接位点)中大约90%位于二级结构的环区或是茎区的端部并靠近环,而且位于环区的G也多靠近环的基部;92%的外显子拼接位点也有类似性质。约82%的分枝点A位于环区或环与茎的连接部位。折叠结构的形成使剪接位点和分枝点在空间上彼此靠近。

蛋白质二组结构由氨基酸和mRNA序列编码

据氨基酸序列，密码子序列和蛋白质二组结构序列的比较，指出约18%的两肽的密码子具有反常的蛋白质结构偏好性，并且这种反常不能用随机涨落解释，因而关于蛋白质折迭的Anfinsen原理可能需要作适当修正。

HSF2 mRNA 在热应激和大剂量11酸睾酮诱导恒河猴生精细胞凋亡中的表达变化

为了探讨 HSF2 mRNA 在热应激和超生理剂量睾酮诱导恒河猴生精细胞凋亡中的表达变化, 作者建立了手术诱导单侧隐睾和注射大剂量11酸睾酮(TU)恒河猴动物模型, 应用3′末端标记分析(TUNEL)和原位杂交方法, 检测睾丸细胞的凋亡信号和 HSF2的表达变化. TUNEL 结果显示热应激和超生理剂量睾酮能够诱导生精细胞出现凋亡信号, 它分别于处理后第5天和第30天达到最强, 表明热应激和睾酮干扰精子发生可能是通过生精细胞凋亡的方式来实现的. HSF2 mRNA 水平在生精细胞凋亡早期(凋亡信号达到最强以前)略有降低, 而在凋亡高峰期之后其表达急剧下降. Hsf2基因与作者以前研究的 Hsp70-2基因的表达具有时间上的相关性, 表明 HSF2蛋白可能调控 Hsp70-2基因的表达, 而且 HSF2可能通过多种方式影响精子的发生以及抑制生精细胞的凋亡。

翻译过程中mRNA的拓扑形态及其链构象分析

对mRNA发夹结构和单链结构的理论模型构建,以及基于对X射线晶体衍射和NMR实验数据的模拟研究显示,在翻译过程中,核糖体上mRNA解折叠后的单链拓扑形态,与mRNA本身的发夹结构密切相关,均呈现为不同程度的螺旋与卷曲相间的构象。通过包括翻译速度在内的调节作用,这种构象可能会影响到新生肽链的折叠。

Is there a genetic relationship between mRNA and the encoded protein at three-dimensional level?

A comparative study on the structures of some mRNAs and their encoded proteins shows an intriguing correlation between the two foldings. Non-random distribution of codons in the secondary structures of mRNAs is also shown, which appears to be in accordance with the conformational properties of amino acids in protein structures to some extent. These results seem to suggest that there may be a kind of genetic relationship between mRNA and protein at three-dimensional level.

Stability and molecular modeling of triplex DNA inhibiting DNA binding protein binding to the core promoter of hepatitis B virus.

Two three-dimensional structure models of the 21nt oligodeoxyribonucleotides, CPI (G3TG-2TGT2G5TG2TGT) and CP3 (TGTG2TGST2GTG2TG3), were constructed by InsightII (MSI) software in IRIS Indigo2 (SGI) workstation using the crystal structure of TAT tripler formation as the template. The initial structures subsequently were minimized by molecular mechanics. The final structures were believed as the dominant conformation. The results showed that the energy of CP1 is lower than that of CP3, and the former is more stable than the latter. Moreover, the results further proved that the 21nt oligodeoxyribo-nucleotide CP1 stably combines with the core promoter (Cp) fragment of hepatitis B virus (HBV) to form a tripler DNA, and CP1 specifically inhibits a specific cellular factor (DNA binding protein) binding to Cp fragment. These results indicated that specific repression of gene transcription of HBV DNA might be possible by tripler-formation DNA.

Expression and regulation of orphan receptor TR2 mRNA in germ cells of cryptorchid testis in rat and rhesus monkey

Expression and cellular localization of orphan receptor TR2 mRNA in relation to germ cell apoptosis in cryptorchid testes of rat and rhesus monkey have been studied by using in situ hybridization and in situ 3'-end labeling of DNA fragments (TUNEL). The results show that: (i) TR2 mRNA is specifically expressed in the germ cells, mainly in the spermatocytes, round and elongated spermatids. The expression level of TR2 mRNA varies with the seminiferous cycle, (ii) In the rat cryptorchid testes on days 3 and 5 after the surgery, the germ cells began to undergo apoptosis with no evident decrease in TR2 mRNA level. On day 7.5, however, most germ cells underwent apoptosis, while the expression level of TR2 mRNA declined markedly, and TR2 mRNA was rarely expressed on day 10 thereafter. (iii) On days 15 and 20 of the cryptorchid testes of rhesus monkey, TR2 mRNA was only expressed in a few of primary spermatocytes and the mRNA was almost undetectable on days 30, 45, 60. These results suggest that TR2 mRNA probably plays an important role in spermatogenesis and germ cell apoptosis.

The topological configuration and conformational analysis of mRNA in translation

The theoretical model construction of mRNA hairpin structure and single-stranded structure as well as the simulation studies on RNA structure determined by the X-ray crystal diffraction and nuclear magnetic resonance revealed that in translation, after mRNA being unfolded into single-stranded structure, its topological configuration was closely correlative with the original hairpin structure. The conformational features of single-stranded mRNA appeared as helical regions alternating with curly regions to different extents, which might exert the influence on the folding of nascent polypeptide by various regulating effects including different translational rates.

Stable RNA hairpins in 88 coding regions of human mRNA

RNA hairpins containing UNCG, GNRA, CUUG (N = A, U, C or G, R = G or A) loops are unusually thermodynamic stable and conserved structures. The structural features of these hairpin loops are very special, and they play very important roles in vivo. They are prevalent in rRNA, catalytic RNA and non-coding mRNA. However, the 5' C(UUCG)G 3' hairpin is not found in the folding structure of 88 human mRNA coding regions. It is also different from rRNA in that there is no preference for certain sequences among tetraloops in these 88 mRNA folding structures.

The identification and quantification of highly stable 'common hairpin' in the dynamic process of co-transcriptional mRNA folding

In our studies, 88 human mRNA samples were collected from the Integrated Sequence-Structure database and then the dynamic process in co-transcriptional mRNA folding was simulated using the RNAstructure version 4.1 program. Through statistical analyses of the frequencies of occurrence of hairpins, a group of special folding structures-the 'common hairpins'-were identified. These 'common hairpins' have lower energies and occur in all the subsequent folding units that formed in the dynamic folding process. By applying the formulas (1)-(4) of the 'common hairpins' statistical model, 163 'common hairpins' were found, to make up about 7% of the total of 2286 hairpins. Classified studies further show that the 'common hairpins' that were studied may oscillate in the dynamic folding process. However, the hairpin loops of the 'common hairpins' and stems proximal to those 'common hairpins' loops maintain topologically stable structures, while other loops and stems distal to the 'common hairpins' loops are shown to be alterable structures. Strikingly, further studies indicate that the stable structures of these 'common hairpins' may have unbeknown effects on controlling the formation of protein structures in the translation process (unpublished results). (c) 2005 Elsevier B.V. All rights reserved.

Exploring consensus mRNA secondary (folding) structure units by stochastic sampling and folding simulation

It is extremely difficult to explore mRNA folding structure by biological experiments. In this report, we use stochastic sampling and folding simulation to test the existence of the stable secondary structural units of-mRNA, look for the folding units, and explore the probabilistic stabilization of the units. Using this method, We made simulations for all possible local optimum secondary structures of a single strand mRNA within a certain range, and searched for the common parts of the secondary structures. The consensus secondary structure units (CSSUs) extracted from the above method are mainly hairpins, with a few single strands. These CSSUs suggest that the mRNA folding units could be relatively stable and could perform specific biological function. The significance of these observations for the mRNA folding problem in general is also discussed. (c) 2004 Elsevier B.V. All rights reserved.