Silent endogenous DNA
Sde4, the Arabidopsis thaliana we discovered before.
Mutation theory shows that the part has lost the luster of transgenic.
This path is dependent in other ways.
RNA-dependent RNA polymerase for six days
(rdr6) (1).sde4 mutant plants also
Defective small interfering RNA (small interfering RNA)
Production and methyl sine inverse element
Atsn 1 (2) is behind the path.
Three days related to rdr2 and Dicer
(dcl3) and other endogenous siRNA (3). it
Probably, this silent path is relevant.
RNA interference (RNAi technology) mediates heterochromatization.
In Schizosaccharomyces cerevisiae
Pombe, which also depends on small interfering RNA, Dicer,
Chapter four. So for investigation.
Two kinds of transgenes: mechanical and inverse elements
Silence, the molecule we study.
The track of Sde4 identity. What we describe here.
How to code the largest subunit 1 in sde4?
Encoding RNA polymerase is different from
One, two and three RNA polymerases in eukaryotic cells
(Its subunit is specified in Arabidopsis thaliana.
Through the prefix nrpa, nrpb or nrpc respectively).
According to the Convention
It's called RNA polymerase, and we will quote it from now on.
This enzyme acts as RNA polymerase IV (oilseed)
(4) The largest subunit code sde4 in the world.
As nrpd 1a。 Sde4 as mentioned earlier.
The mutation is now designated as nrpd 1a- 1.
Sainsbury Laboratory, Norich John Building Center, 1.
Nr4 7uh, UK. Norich east of england University.
British Nr4 7tj.
* To whom the communication should be sent.
E-mail: David.baulcombe @ it's Sainsbury-laboratory.ac.uk.
Mutants with defects on the screen, across
Arabidopsis strains are used for gene silencing.
(gxa), in which green fluorescent protein
(GFP) transgenic (Figure s 1a, named G)
Silence, by potato X virus (pvx)- green fluorescent protein.
Transgenic (figure s 1a, name 1) (1). unlikeness
Rdr6 mutant, in which the green fluorescent protein is completely lost.
Silence in gxa background, nrpd 1a- 1
Shows a picture of a delayed attack in silence
Plants at the growing point (figure 1A and figure.
S 1f) (1). In seedlings, in the early stage of leaves.
The fluorescence of green fluorescent protein appeared, and within a week,
Silence appears in local areas, and then
Scattered all over the leaf. This delay
Phenotype lasts until flowering.
The young inflorescences are all GFP fluorescent.
The transgenic mRNA of this model.
Small molecules will interfere with RNA accumulation accordingly.
Fluorescence quantity of green fluorescent protein. Therefore, North
Analysis of wild type and nrpd 1a- 1 flower
This indicates that the accumulated GFP has increased.
Expression of mRNA (4.5 times) and RNA of pvx- green fluorescent protein gene.
Nrpd 1a- 1 corresponds to six-fold reduction.
Green fluorescent protein siRNA of 2 1-24-day-nucleotide (nt) (fig.
1B) relative wild type. Complete silence
Rose leaves, where nrpd 1a mediates losses.
Silence is not significantly higher than.
Amount of flowers, green fluorescent protein gene and siRNA
Comparable.
RNA-mediated DNA methylation (rddm)
Is a silent plant, and
A possible small molecule interferes with RNA guidance.
Complicated. Consistent rddm in green fluorescent protein
In gxa plants, the DNA methylation trajectory of green fluorescent protein gene is
Rdr6 and sgs3 mutants were lost in flowers.
23 。 Rompuy Hoff et al., Williams Year. Mediterranean. 197 , 1525 ( 2003 ) 。
24 。 Spoon egawa et al., at present. biology 13 , 1252 ( 2003 ) 。
25 。 Supported by the national medical service system (r37-ai33443). we think
Thank you, J. Bob, ten grandchildren per hour, and D. rostain.
Read this manuscript carefully and the Tenth Forest (University)
Buffalo), in order to share the defects of card 1 1.
Jurkat cells. Molecular interaction data have been
Stored in the biomolecular interaction network
Database and login codes 209039 to 209044.
Support online materials
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Wireless TTL flash
Materials and methods
I to III in the picture
165438+20041October 4th; Accepted, June 65438+10/October 65438+April 2005
10. 1 126/ science. 1 107 107
SiRNA with green fluorescent protein (1) (Figure 1 C). In nrpd 1a- 1
The siRNA of flowers, such as green fluorescent protein, decreased, but
There was no absence, we only found a slight change.
DNA methylation pattern of green fluorescent protein (figure 1 C)
This contrast is a more obvious loss.
Atsn 1 DNA methylation nrpd 1a- 1 plants
( 2 , 5 ) 。 So, the nrpd 1a channel, right?
The only suitable siRNA is rddm.
At the beginning, nrpd 1a- 1 was quite rearranged.
Chromosome 1 disorder
At 1g63020 (figure 1, 2 to 7), and then use.
Other nrpd 1a alleles are associated with.
Confirm with nrpd1a.
The gene encodes nrpd1a. sequence
Non-alignment of nrpd 1a and Arabidopsis nrpd 1a
Homologous (code at2g40030) and two
Protein is unique to rice.
RNA branching into the largest subunit
Polymerase (figs. 2A and s2, a to c).
Consistent nrpd 1a is the largest in the world.
Subunit-Duoya unit oil four, rice
The Arabidopsis genome encodes two.
Protein can be the second largest.
Subunits (at3g 18090 and at3g23780) (6)
(figure 2B and figure s2d). Test mute
The role of these genes in Arabidopsis nrpd2
Subunit, we transform wild-type gxa.
Construction of plant reverse repeat sequence
Nrpd 1a targets RNAi, which is suspected.
Nrpd2 gene, or Adan (as control group) (Figure 3,
A and b). Conversion and infrared
For nrpd 1a and suspected
Nrpd2 gene, but not for Adan (Figure 3, C
And d) delayed onset of silence.
Such nrpd 1a- 1. Although two nrpd2
Genes will be inactivated and constructed by infrared rays.
(Figure s3b), the evidence of two routes shows that,
Active nrpd2 trajectory, not in 3g23780.
Compared with at3g 18090. First, gene-specific reversal
Transcription polymerase chain reaction (RT-PCR)
The analysis results show that most nrpd2
From at3g23780 (figure s3e)
Secondly, a mutant at3g23780 was inserted.
(nrpd2a- 1, figure s3f) instead of at3g 18090.
(nrpd2b- 1, figure s3f) eliminates the internal source.
Small molecules interfere with RNA (fig. 3A).
Nrpd 1a and nrpd2 have similarities and differences in order.
They nrpa, nrpb, nrpc
Homologues, corresponding functions in this region
Important characteristics of yeast RNA polymerase
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report
Figure. 1 。 GFP silences nrpd1a-(1)1. Ultraviolet photo gxa wild
Type (WT) and nrpd 1a- 1 flowering plants. (2) northern analysis of green fluorescent protein
The expression of mRNA and siRNA in flowers (VI), stems (S) and leaves (1) consists of WT and SiRNA.
Nrpd 1a- 1 gxa factory. (3) Southern blot analysis and purification of genomic DNA.
Two (7). For nrpd 1a, region and identity.
The Nrpb 1 includes an n-terminal clamping core.
(20%), zinc metal binding site, activity.
Website (4 1%), funnel and some cracks.
Domain (initial, 42%, final elimination, 24%). should
Fault domain in nrpa 1
Nrpb 1 and nrpc 1 are missing in nrpd1a.
(conservative area 7) (fig. s2b). however
C-end clamp core (Figure s2b), which defines
Before the end of the spherical domain
C-terminal domain (CTD is) nrpb 1 (7), is
Currently (23% same). Nrpd2 is 34% the same.
There are nrpb2 and conservative areas.
The corresponding binding sites are very small.
Core subunits (nrpb3/ac40, nrpb 10,
Nrpb 12) (Figure s2d), which is how the train of thought works.
Act on that enzyme assembly (7). how heavy
Gene nrpb3/ac40, some enterprises
They can know the details of Petroleum IV. However, for
There are only two genes in nrpb 10 and nrpb 12.
Everyone in Arabidopsis thaliana, they can share.
Between the fourth oil seed and other RNA polymerases.
Because they are in other eukaryotes (8).
The most striking difference
Nrpb 1 and nrpd 1a are located at the c-terminal.
Nrpd 1, such as rice and Arabidopsis thaliana.
Protein has a similar C-terminal.
Half an uncoded protein (defective
Chloroplasts and leaves) regulate the S rRNA gene.
Processing chloroplasts (9) (Figure s2a). this
C-terminal extension, coordinating RNA
Polymerase activity and downstream processing
Step to be similar to.
CTD belongs to polii (8). Combination book
Unusual C-terminal and conservative features
Sense RNA polymerase showed that Polish IV was.
Active RNA polymerase and its important differences
One, two and three from sense RNA polymerase.
From the same tissue, such as item (2), digestion and methylation sensitivity.
It was found that the enzyme sau96i was GFP. Unmethylated g and gxa sgs3 systems
As a control. The DNA fragment is greater than NT$ 554 because of DNA.
Methylation.
Figure. 2 。 phylogenetic tree
Maximum (a) and
Second largest RNA
Polymerase subunit family
From plants, worms and
Yeast. right/right-hand side
Show RNA to each tree
Polymerase gray subfamily
And a black box display.
Four subunits of Arabidopsis oil.
In order to further explore the mechanism of oil production.
Four interfaces are silent, and another feature of us is that
Mutation and GFP delayed silencing
(figure s4a). It has an inverted chromosome 4.
This will destroy rdr2 (Figure s4b, rdr2-3), and
It can supplement the silent phenotype of green fluorescent protein gene.
By transformation with wild-type rdr2
Genomic DNA (fig. s4a). Variation here, because in
Every nrpd 1a mutation of nrpd2a- 1 and the mutation with
Get a lower amount of endogenous NT$ 24 yuan.
SiRNA (i.e. atsn 1, 1003, 02 and group 2), and
In the wild type, and mir 167 amount.
Not affected at (3) (Figure 3A). These siRNAs are all
Taiwan dollars around 24 yuan, dcl3 Dicer is
Students involved (3). It's possible.
Therefore, the oil four, rdr2 and dcl3 methods.
Unite in a silent way. Four generals of petroleum
The generated RNA species are copied to
Double stranded RNA (double stranded RNA) rdr2.
This double-stranded RNA is then processed into
Small molecules interfere with RNA through dcl3.
The mutations of rdr2 and rdp 1
Through long-term RNA, small molecules interfere with RNA gene sites.
However, the opposite effect on small molecules will interfere with RNA,
Long-term RNA is richer in silence.
Mutant. Therefore, there are more atsn 1 RNA.
Rich in nrpd 1a and rdr2 mutants.
In wild-type plants (fig. 3 C), it is shown that
They are not oil report cards. Presumably, this
The result of RNA must be determined by RNA polymerase III.
Transcription (10) atsn 1 is silence.
Tetra-rdr2-dcl3 pathway of oil plants is wild type.
Plants. We can't detect it, just oil four.
Atsn 1 Presumably this is because.
They are currently few in number.
Is produced by more abundant RNA that is masked.
Other RNA polymerase.
The 4-rdr 2-DC L3 pathway of this oil is related.
Unlike group 2 and siRNAs in 2002, it is not necessary.
Ago4 or DNA methylation (3). however
Atsn 1 and 1003 are highly methylated.
Among wild-type plants, nrpd 1a is 10000 mu.
Www.sciencemag.org Science Volume 308 April 2005 1
report
Figure. 3 。 Molecular indicator silence. (a) analysis of the position of the north on the 9th and 9th; 10 lane nrpd 1a-3 lane 1 1, nrpd1a-4; 12 lane, nrpd2a- 1 lane 13 lane.
Endogenous MicroRNA: Lane 1 to 8, gxa (C24) background; Track 9 to nrpd2b- 1. Stains have been removed and repainted many times. (2) South District
13, lieutenant colonel background -O. 1 line, per gram; Three repeats, four repeats and two repeats of Lane 2, rdr2-3 and Wuchang rDNA were digested by a series of alleles of nrpd 1a and compared with other alleles.
Independent RDR2-3 RDR2P:: RDR2T2 qualification line; 5 lanes, nrpd1a-1; Lane 6 and 7, methylation sensitive enzyme hpaii. (c) Endogenous reverse transcription polymerase chain reaction analysis.
Two independent nrpd1a-1nrpd1AP:: nrpd1t 2 qualification lines; 8 lane nrpd1a-2; Atsn 1 is an elevated mutant of rdr2-2 and nrpd1a.
Tants (Figure 3B), accumulate yourself.
SiRNAs depends on argonaute protein.
Ago4, de novo DNA methyltransferase
Drm 1 and drm2 (No.5, 1 1 and 12), and oil.
Fourth, rdr2, dcl3. In these examples, it
It seems that we have proposed it in the first Pompeii.
(13), that is, keeping silent involves
Self-reinforcing silence path is among them.
The production of siRNA mediated by oilseed four is dependent.
About ago4-mediated DNA methylation
or vice versa, Dallas to the auditorium
Here you can, as described in volume 4.
Solving a Paradox Chromatin Silence
This mechanism depends on RNA. if
Silent gene transcription consists of a single
Polymerase, this RNA-dependent mechanism
Will not be stable because of depression.
Transcription from these sites
It can also lead to the loss of silent RNA.
However, specific polymerase images are silenced.
Oil four, can resist chromatin or DNA.
Modification affects polymerase one to three.
By analogy, you can remain silent steadily.
State. Four branches of oil in animals and fungi
Absence, but the paradox of silence may
Solved, if there is a form of RNA.
Polymerase 1-3 and silencing specificity
A subunit that makes transcribed heterochromatin.
Therefore, maintenance
RNA-dependent silencing.
Finally, the general silence.
Its mechanism is GFP crossover.
Gene silencing and endogenous siRNA
The road to silence is dependence.
For * * * and connexin (figure 10 18). In the rose leaves
These two silent paths of plants
Are independent of each other, because
Gene silencing is not affected by rdr2 (fig. 10 18).
Due to the persistence of endogenous 24nt siRNA
Plants in rdr6 (2). However, among these flowers,
Because of the green fluorescent protein.
Silence is influenced by two rdr2 and rdr6.
This potential silent way of interaction,
Combined with one's own ability to form
Feedback and self-reinforcement cycle (14), explanation
Endogenous potential complexity
SiRNAs involved in regulating the network.
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Support online materials
www . science mag . org/CGI/content/full/ 1 1069 10/DC 1
Materials and methods
Figure Table 1 to Table 4
Tables S 1 and S2
References and bonds
2004129 October 10; Accepted, 65438+2005125 October
Published on the Internet on February 3, 2005;
10. 1 126/ science. 1 1069 10
Include this information, please refer to this document.
Translation operator gene
For ribosomes: how to inhibit substances
Protein ruled out ribosome binding.
Lars Jenner, 1 Pascal Longby, 2 Bernard Rees, 1
Clemens Schulz-Bliss, the third student, springer, 4 chantal Ehresmann, 2.
Bernard Ehresmann, Dino maurras, 1 Gulnara Yusupova, 1.
Yusupov 1，2 *
Thermophilic ribosomes were transferred from cocrystallization to sponsors.
Rna (tRNA molecule) and intact messenger RNA (mRNA) carry translation.
Operator. Road mRNA is defined with a resolution of 5.5 angstroms.
More complex than ribosomes with the same crystal structure.
There was no deletion or simultaneous expression of structural mRNA. Precise ribosome environment
The stem ring of the post operator is perpendicular to the surface.
The11930s subunit of ribosomes on this platform. Binding operators and
The initiator's tRNA appears at the site where ribosomes and an uninhabited tRNA are evacuated.
It is expected to be complicated at first. Positioning supervision
The operator of this domain clarified the molecules relative to ribosomes.
Mechanism, that is, constraints inhibit the over-translation of sub-switches. Our data
Suggest how to express control elements in general.
Ribosomes perform their regulatory tasks.
When the initial translation is generally accepted, it needs rapid adaptation. conventional
Efficiency promotes the synthesis of protein, and protein is several elements involved in ribosomes of eubacteria.
The key step is to control the expression of genes and the dynamics of gene influence research.
April 2005 1 Volume I 308 www.sciencemag.org Science
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