scholarly journals TPR domain coding gene ST2 may be involved in regulating tillering and fertility in rice

2021 ◽  
Author(s):  
gang wei ◽  
hongxia yang ◽  
zixian xiong ◽  
jingwen wu ◽  
danyang chen ◽  
...  
Keyword(s):  
Male Sterility ◽  
Nuclear Gene ◽  
Tiller Number ◽  
Pcr Analysis ◽  
Tetratricopeptide Repeat ◽  
Subcellular Localisation ◽  
Initial Stage ◽  
Ubisch Body ◽  
Evolutionarily Conserved ◽  
Tpr Domain

A decrease in the tiller number and male sterility will lead to a decline in the rice yield. Therefore, it is significant to study the molecular mechanism of controlling the tiller number and regulating the male reproductive development. The mutant st2 (single tiller 2) was induced by ethyl methane sulfonate (EMS) in the indica maintainer line Xinong 1B and showed single tillering and male sterility. I<sub>2</sub>-KI staining showed that the st2 pollen was aborted. The scanning electron microscope (SEM) observation underlined that the anther of st2 became smaller, the wax of the epidermis reduced, the inner wall shrank and the Ubisch body decreased, the pollen collapsed, and the germination pore developed abnormally. The genetic analysis discovered that the trait was controlled by a single recessive nuclear gene located on chromosome 3. LOC_Os03g05540 encoding a tetratricopeptide repeat (TPR) domain was identified as the candidate gene by sequencing. The quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that ST2 was highly expressed in the stem apical meristem (SAM) and the initial stage of meiosis during the anther development. The subcellular localisation indicated that ST2 is a nuclear and plasmic localisation protein. The homology analysis demonstrated that ST2 was evolutionarily conserved. These results laid a foundation for further study of the ST2 function.

scholarly journals Dissection of Swa2p/Auxilin Domain Requirements for Cochaperoning Hsp70 Clathrin-uncoating Activity In Vivo

2006 ◽  
Vol 17 (7) ◽  
pp. 3281-3290 ◽  
Author(s):  
Jing Xiao ◽  
Leslie S. Kim ◽  
Todd R. Graham
Keyword(s):  
Atpase Activity ◽  
Point Mutation ◽  
Mutational Analysis ◽  
Weak Interactions ◽  
Tetratricopeptide Repeat ◽  
Uba Domain ◽  
J Domain ◽  
Tpr Domain

The auxilin family of J-domain proteins load Hsp70 onto clathrin-coated vesicles (CCVs) to drive uncoating. In vitro, auxilin function requires its ability to bind clathrin and stimulate Hsp70 ATPase activity via its J-domain. To test these requirements in vivo, we performed a mutational analysis of Swa2p, the yeast auxilin ortholog. Swa2p is a modular protein with three N-terminal clathrin-binding (CB) motifs, a ubiquitin association (UBA) domain, a tetratricopeptide repeat (TPR) domain, and a C-terminal J-domain. In vitro, clathrin binding is mediated by multiple weak interactions, but a Swa2p truncation lacking two CB motifs and the UBA domain retains nearly full function in vivo. Deletion of all CB motifs strongly abrogates clathrin disassembly but does not eliminate Swa2p function in vivo. Surprisingly, mutation of the invariant HPD motif within the J-domain to AAA only partially affects Swa2p function. Similarly, a TPR point mutation (G388R) causes a modest phenotype. However, Swa2p function is abolished when these TPR and J mutations are combined. The TPR and J-domains are not functionally redundant because deletion of either domain renders Swa2p nonfunctional. These data suggest that the TPR and J-domains collaborate in a bipartite interaction with Hsp70 to regulate its activity in clathrin disassembly.

Cloning and function analysis of BAG family genes in wheat

2016 ◽  
Vol 43 (5) ◽  
pp. 393
Author(s):  
Shiming Ge ◽  
Zhen Kang ◽  
Ying Li ◽  
Fuzhen Zhang ◽  
Yinzhu Shen ◽  
...  
Keyword(s):  
Heat Stress ◽  
Salinity Stress ◽  
Heat Tolerance ◽  
Expressed Sequence Tag ◽  
Function Analysis ◽  
Pcr Analysis ◽  
Subcellular Localisation ◽  
Hybrid Technique ◽  
Rt Pcr ◽  
Conserved Domain

By analysing the cDNA microarray of the salt tolerant mutant of wheat RH8706–49 under salinity stress, our results showed an expressed sequence tag fragment and acquired an unknown gene (designated as TaBAG) with a BAG conserved domain through electronic cloning and RT–PCR technology. The gene was registered into GenBank (No. FJ599765). After homologous alignment analysis, electronic cloning, and amplifying with RT–PCR, the other gene with a BAG conserved domain, TaBAG2, was obtained and registered into GenBank (No. GU471210). Quantitative PCR analysis demonstrated that TaBAG2 expression was induced by saline and heat stress. TaBAG gene expression under salinity stress increased remarkably but showed an insignificant response to heat stress. The adversity stress detection results showed that Arabidopsis overexpressing TaBAG and TaBAG2 exhibited an obvious salt tolerance increase. Under heat stress, Arabidopsis overexpressing TaBAG2 showed increased heat tolerance; however, the heat tolerance of Arabidopsis overexpressing TaBAG did not vary significantly under heat stress. Subcellular localisation results demonstrated that TaBAGs were mainly located in the cytoplasm and the cell nucleus. We applied fluorescence complementation and yeast two-hybrid technique to prove that TaBAG2 can obviously bond with TaHsp70 and TaCaMs. After the respective mutation of aspartic acid (D) and arginine (R) at high conservation in BAG domain of TaBAG2, the bonding interaction between TaBAG2 and TaHsp70 disappeared, indicating that the two amino acids were the key loci for the interaction between TaBAG2 and TaHsp70. Heat tolerance detection results demonstrated that the heat tolerance of Arabidopsis overexpressing and cotransfected with TaBAG2 and TaHsp70 was much higher than that of Arabidopsis overexpressing TaBAG2 and Arabidopsis overexpressing TaHSP70. This finding implies that the synergistic use of TaBAG2 and TaHSP70 can improve heat tolerance of plants.

Combined transcriptome sequencing and prokaryotic expression to investigate the key enzyme in the 2-C-methylerythritol-4-phosphate pathway of Osmanthus fragrans

2020 ◽  
Vol 47 (10) ◽  
pp. 945
Author(s):  
Rui Xiong ◽  
Zhu Chen ◽  
Weiyu Wang ◽  
Li Jiang ◽  
Yan Xiang ◽  
...  
Keyword(s):  
Prokaryotic Expression ◽  
Expression Vectors ◽  
Pcr Analysis ◽  
Subcellular Localisation ◽  
Reaction Products ◽  
Osmanthus Fragrans ◽  
Qrt Pcr ◽  
Key Enzyme ◽  
Series Of Experiments ◽  
Main Components

Terpenoids are one of the main components of plant aromas. In the present study we investigated these compounds in Osmanthus fragrans Lour., which is a fragrant plant widely used for the production of essential oils. Quantitative real-time PCR (qRT-PCR) results of enzymes associated with the 2-C-methylerythritol-4-phosphate pathway confirmed that the TPS is a key enzyme for terpenoid synthesis in O. fragrans. In a series of experiments, we identified the TPS candidate genes in O. fragrans and revealed the underlying catalytic activity and subcellular localisation of the encoded proteins. Because there is no available O. fragrans reference genome, we sequenced and analysed its transcriptome and identified two putative TPS genes, OfTPS1 and OfTPS2. According to qRT-PCR analysis, both genes were most highly expressed at the full-bloom stage, suggesting that OfTPS1 and OfTPS2 contribute to O. fragrans terpenoid synthesis. To verify this hypothesis, we constructed prokaryotic expression vectors to obtain protein. In order to study the function of OfTPS1 and OfTPS2 in the synthesis of monoterpenes, the obtained proteins were reacted with geranyl pyrophosphate. As a result, two kinds of monoterpenes, (E)-β-ocimene and linalool, were detected from reaction products, respectively. In conclusion, OfTPS1 and OfTPS2 are both monoterpene synthases.

scholarly journals Cytological observation of anther structure and genetic investigation of a thermo-sensitive genic male sterile line 373S in Brassica napus L

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Yanyan Sun ◽  
Dongsuo Zhang ◽  
Zhenzhen Wang ◽  
Yuan Guo ◽  
Xiaomin Sun ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Male Sterility ◽  
Spontaneous Mutation ◽  
Hybrid Breeding ◽  
Pcr Analysis ◽  
Cytological Observation ◽  
Male Sterile ◽  
Pollen Abortion ◽  
Genetic Investigation ◽  
Temperature Changes

Abstract Background Photoperiod and/or thermo-sensitive male sterility is an effective pollination control system in crop two-line hybrid breeding. We previously discovered the spontaneous mutation of a partially male sterile plant and developed a thermo-sensitive genic male sterile (TGMS) line 373S in Brassica napus L. The present study characterized this TGMS line through cytological observation, photoperiod/ temperature treatments, and genetic investigation. Results Microscopic observation revealed that the condensed cytoplasm and irregular exine of microspores and the abnormal degradation of tapetum are related to pollen abortion. Different temperature and photoperiod treatments in field and growth cabinet conditions indicated that the fertility alteration of 373S was mainly caused by temperature changes. The effects of photoperiod and interaction between temperature and photoperiod were insignificant. The critical temperature leading to fertility alteration ranged from 10 °C (15 °C/5 °C) to 12 °C (17 °C/7 °C), and the temperature-responding stage was coincident with anther development from pollen mother cell formation to meiosis stages. Genetic analysis indicated that the TGMS trait in 373S was controlled by one pair of genes, with male sterility as the recessive. Multiplex PCR analysis revealed that the cytoplasm of 373S is pol type. Conclusions Our study suggested that the 373S line in B. napus has a novel thermo-sensitive gene Bnmst1 in Pol CMS cytoplasm background, and its fertility alteration is mainly caused by temperature changes. Our results will broaden the TGMS resources and lay the foundation for two-line hybrid breeding in B. napus.

Analysis of mitochondrial DNA, chloroplast DNA, and double-stranded RNA in fertile and cytoplasmic male-sterile sunflower (Helianthus annuus)

1986 ◽  
Vol 28 (1) ◽  
pp. 121-129 ◽  
Author(s):  
Gregory G. Brown ◽  
Howard Bussey ◽  
Lee J. DesRosiers
Keyword(s):  
Male Sterility ◽  
Restriction Analysis ◽  
Nuclear Gene ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Male Sterile ◽  
Gene Effects ◽  
Mitochondrial Dnas ◽  
Specific Association ◽  
Chloroplast Dnas

The extent of variation in the mitochondrial DNAs (mtDNAs), chloroplast DNAs (ctDNAs), and double-stranded RNAs (dsRNAs) of sunflower lines carrying fertile and male-sterility conferring cytoplasms was examined. To minimize nuclear gene effects, efforts were concentrated on two chromosomally isogenic lines, CM400 (fertile) and cmsCM400 (male sterile), which differ only in their cytogenes. A circular 1.45 kilobases (kb) plasmid DNA was found in the mitochondria of the four fertile lines examined, but was absent in the male-sterile line. Restriction enzyme analysis of mtDNAs of the fertile and male-sterile cytoplasms with BamHI, EcoRI, and HindIII revealed no fragment mobility differences between them other than those which could be ascribed to the 1.45-kb circle. Similar restriction analysis of ctDNA showed no differences between fertile and male-sterile cytoplasms. Both CM400 and cmsCM400 contain dsRNA molecules. The number and sizes of these dsRNAs varied from preparation to preparation in both lines. Species of 3.3 and 1.5 kb, which were the only dsRNAs common to all preparations from CM400, were also the only species common to all preparations from cmsCM400. Thus, no consistent differences between the fertile and male-sterile cytoplasms were seen in these molecules. The specific association of the 1.45-kb plasmid with fertile cytoplasm together with the absence of variation in ctDNA and dsRNA, suggests the involvement of mtDNA in sunflower cytoplasmic male sterility.Key words: DNA (mitochondrial), sterility (male), sterility (cytoplasmic), Helianthus, sunflower, DNA chloroplast.

scholarly journals Knockout of SlMS10 Gene (Solyc02g079810) Encoding bHLH Transcription Factor Using CRISPR/Cas9 System Confers Male Sterility Phenotype in Tomato

Plants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1189
Author(s):  
Yu Jin Jung ◽  
Dong Hyun Kim ◽  
Hyo Ju Lee ◽  
Ki Hong Nam ◽  
Sangsu Bae ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Male Sterility ◽  
Mutant Line ◽  
Pcr Analysis ◽  
Bhlh Transcription Factor ◽  
Hybrid Seed Production ◽  
Sequencing Analysis ◽  
Alternative Approach ◽  
Deep Sequencing Analysis ◽  
Male Sterility Phenotype

The utilization of male sterility into hybrid seed production reduces its cost and ensures high purity of tomato varieties because it does not produce pollen and has exserted stigmas. Here, we report on the generation of gene edited lines into male sterility phenotype by knockout of SlMS10 gene (Solyc02g079810) encoding the bHLH transcription factor that regulates meiosis and cell death of the tapetum during microsporogenesis in the tomato. Twenty-eight gene edited lines out of 60 transgenic plants were selected. Of these, eleven different mutation types at the target site of the SlMS10 gene were selected through deep sequencing analysis. These mutations were confirmed to be transmitted to subsequent generations. The null lines without the transferred DNA (T-DNA) were obtained by segregation in the T1 and T2 generations. In addition, we showed that the cr-ms10-1-4 mutant line exhibited dysfunctional meiosis and abnormal tapetum during flower development, resulting in no pollen production. RT-PCR analysis showed that the most genes associated with pollen and tapetum development in tomatoes had lower expression in the cr-ms10-1-4 mutant line compared to wild type. We demonstrate that modification of the SlMS10 gene via CRISPR/Cas9-mediated genome editing results in male sterility of tomato plants. Our results suggest an alternative approach to generating male sterility in crops.

scholarly journals Tcc1p, a Novel Protein Containing the Tetratricopeptide Repeat Motif, Interacts with Tup1p To Regulate Morphological Transition and Virulence in Candida albicans

2006 ◽  
Vol 5 (11) ◽  
pp. 1894-1905 ◽  
Author(s):  
Aki Kaneko ◽  
Takashi Umeyama ◽  
Yuki Utena-Abe ◽  
Satoshi Yamagoe ◽  
Masakazu Niimi ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Fungal Pathogen ◽  
Affinity Purification ◽  
Specific Protein ◽  
Tetratricopeptide Repeat ◽  
Human Fungal Pathogen ◽  
The Common ◽  
Transcriptional Gene Regulation ◽  
Tpr Domain ◽  
Novel Protein

ABSTRACT The transcriptional factor CaTup1p represses many genes involved in intracellular processes, including the yeast-hypha transition, in the human fungal pathogen Candida albicans. Using tandem affinity purification technology, we identified a novel protein that interacts with CaTup1p, named Tcc1p (Tup1p complex component). Tcc1p is a C. albicans-specific protein with a 736-amino-acid polypeptide with four tetratricopeptide repeat (TPR) motifs in the N-terminal portion. Tcc1p formed a protein complex with CaTup1p via the TPR domain of Tcc1p, independently of CaSsn6p-CaTup1p The tcc1Δ disruptant showed filamentous growth under conditions inducing the yeast form, as is true of the Catup1Δ mutant. Consistent with this result, the common set of hypha-specific genes was negatively regulated by both TCC1 and CaTUP1. These observations will provide new insights into CaTup1p-dependent transcriptional gene regulation in C. albicans.

scholarly journals Human U4/U6 snRNP Recycling Factor p110: Mutational Analysis Reveals the Function of the Tetratricopeptide Repeat Domain in Recycling

2004 ◽  
Vol 24 (17) ◽  
pp. 7392-7401 ◽  
Author(s):  
Jan Medenbach ◽  
Silke Schreiner ◽  
Sunbin Liu ◽  
Reinhard Lührmann ◽  
Albrecht Bindereif
Keyword(s):  
Amino Acids ◽  
Protein Interactions ◽  
Rna Binding ◽  
Mutational Analysis ◽  
Protein A ◽  
Tetratricopeptide Repeat ◽  
Terminal Sequence ◽  
Recognition Motifs ◽  
Tpr Domain

ABSTRACT After each spliceosome cycle, the U4 and U6 snRNAs are released separately and are recycled to the functional U4/U6 snRNP, requiring in the mammalian system the U6-specific RNA binding protein p110 (SART3). Its domain structure is made up of an extensive N-terminal domain with at least seven tetratricopeptide repeat (TPR) motifs, followed by two RNA recognition motifs (RRMs) and a highly conserved C-terminal sequence of 10 amino acids. Here we demonstrate under in vitro recycling conditions that U6-p110 is an essential splicing factor. Recycling activity requires both the RRMs and the TPR domain but not the highly conserved C-terminal sequence. For U6-specific RNA binding, the two RRMs with some flanking regions are sufficient. Yeast two-hybrid assays reveal that p110 interacts through its TPR domain with the U4/U6-specific 90K protein, indicating a specific role of the TPR domain in spliceosome recycling. On the 90K protein, a short internal region (amino acids 416 to 550) suffices for the interaction with p110. Together, these data suggest a model whereby p110 brings together U4 and U6 snRNAs through both RNA-protein and protein-protein interactions.

Sign in / Sign up

Export Citation Format

Share Document