ks06 - nuclear Plasma 9n “slope membrane SPLICING R NA...

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Unformatted text preview: nuclear Plasma 9n “slope membrane SPLICING R NA TRANSPOR T hQ 5V— THANSLATION 0F “MSW” MESSENGER mun b v-v protein -1... v var/E Figure 8—2 Protein synthesis {DNA -> RNA —r protein) in eucaryotes. Eucaryotic cells have evolved numerous membrane- hounded compartments that segregate their various chemical reactions so as to make them more efficient. and the nucleus is one such compartment. The nuclear envelope keeps functional ribosomes out ofthe nucleus. preventing RNA transaipts from being translated into protein until they have been extensively processed (spliced) and transported out of the nucleus into the cytosol. 'l‘hns RNA splicing and transport steps are interposed between DNA transcription and RNA translation. _ aeell _ __ _ ocell 1/ [Ill/15’ ' MCMI m use m l P’ 0 ‘1 | | i | | i | | P | l \. _.__ _. . K... . _ Figure 3 Transcriptional control of o-specific and a—specific genes by Matulp, MataZp, and Mcmlp. Appropriate expression of a-specific and a-spccific genes in an a cell (lefl) and in an a cell (right) is mediated by interactions of Matolp and Mata2p with Mcmlp in the upstream regulatory regions of the targeted genes. (P) Mcmlp-binding site; (0) Matalp-binding site. The P sites are drawn differently for o-spccific genes and for Il-spccific genes to indicate that Mcmlp does not bind in the same way to both (see text). In an a cell, Mcmlp is able to activate transcription of a-specific genes. In an a cell, Matolp and Mcrnlp assist each other's binding. which allows transcrip- tion of the a-specific genes. In anotcell, MataZp and Mcmlp assist each other‘s binding, which results in repression of the a-specific genes. RNA E Figure 18-20 A schematic yeast gene showing the relative positions of its control ele- ‘I }—-——i merits. I m-mbp l——————i : 20—40%.) I —— __ i ' sa—soobp if. L ,-.-Harvrc' \ r... \\ flJr-J HF, \ H (0112) 3‘ untrunskltted region oding “quanta {_Afi RNA slop Figure 8—51 The transcribed portion of the human B-globln gene. The sequence of the DNA strand corresponding to the mRNA sequence is given, with the primary RNA transcript surrounded by a green line and the nucleo- tides in the three exons shaded yellow. Note that exon 1 includes a 5’-ieader sequence and that exon 3 includes a 3’-untranslated sequence; although these sequences are included in the mRNA, they do not code for amino acids. The highly conserved GT and AG nucleotides at the ends of each intron are boxed [see Figure 8—53]. along with the cleavage and poly- adenylation signal near the 3' end of the gene (MTMA. see Figure H9). binding target pa rt ner protein transcriptional IONA-binding domain activation domain BA|T PREY RECOMBINANT GENES ENCODING BAIT AND PREY INTRODUCED INTO yeast cell YEAST CE LL CAPTURED PREY TRANSCRIPTION OF REPORTER — reporter protei transcriptional activator NE binding site Figure— The yeast two-hybrid system for detecting protein-protein interactions. The target protein is fused to a DNA-binding domain that localizes it to the regulatory region of a reporter gene as When this target protein binds to another specially designed protein in the cell nucleus ("pref"). their interaction brings together two halves of a transcriptional activator. which then switches on die expression of the reporter gene.The reporter gene is often one that will permit growth on a selective medium. Bait and prey fusion proteins are generated by standard recombinant DNA ted'miques. In most cases. a single bait protein is used to fish for interacting partners among a large collection at prey proteins produced by ligating DNA encoding the activation domain of a transcriptional activator to a large mixture of DNA fragments from a cDNA library. atom-rm. acacccmcccncoccn Atcrscmccaocaccacrmcsccatmc comm Tmotcnsoocmnc 5' GCAC'I GACTCTC'ICTGCCTA‘I'IGGTCTAT TTCCCAOCCT E ‘GTCTA‘lGGGACCCITGATGTTI TC'I “CC CCTTCTT‘I‘TCTATGGT‘IMG'I‘TCATGTCAT AWTMCAGGGTW TTCTCCGCMT‘ITTI‘ACTATTA‘E‘ACTTAA TCXDCTTMCATTG‘IB’E‘ATAACAAMGGAM ATC'DCTGAGATACA'ITARG TWITAAM acacw'rc'tt'macmcan - mart-accura'rntcmrccnsrt'mc camraarcrccc'mc'mmrrrcrr N amnmrrcaracaTM-rmnatsc c mma’rcoot‘rmcaomsrorrrrm 9 ammacacamrtctccmmcccs E mnmrmmnnmmcm rmmmamamcrnrrrctrmtc TTRTT'TCTMTWMECTCTTTC rmaccocaaraamxmcaaacmscm GCCTCTTTGCACCATICIWGAATWG tcarmrncmccflmoocanmocmr armcrccamrmramcrccamrss ATTGTAAC‘IGATGTMGAGGTTTCATRTTG craarltccaoc'rm-rocssc-rnccarrc TGCTT‘I‘TAT‘I'I‘TAWTMGGCTG exon 3 TGATGTATI‘TAM‘PTAT’ITCTGIQTATTH 3‘ ACTAMMGGGAANWTGCA TTTAMACATAMGAAATGATGKGCTGTW WWAMATACACTATAT‘CTTW CTOCATWGMDGTGAGGCTGCAACCAG CTMWTTGGCAACAWCCCTGATGC CTATGCC'ITATTCATCCCTCAGMMGGAT TCWGTAGAGGCT’i‘G-R‘I'TTGCRGGTTAMG TT'ITGCTAMTTGTATT'TTACATI‘RCTTAT TGTN'I‘AGCTGTCCTCATGM'I‘GNTTTTC ...
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