ARRANGEMENT OF MESSENGER-RNA AT THE DECODING SITE OF HUMAN RIBOSOMES - 18S-RIBOSOMAL RNA NUCLEOTIDES AND RIBOSOMAL-PROTEINS CROSS-LINKED TO OLIGOURIDYLATE DERIVATIVES WITH ALKYLATING GROUPS AT EITHER THE 3' OR THE 5' TERMINI

被引:49
作者
MALYGIN, AA
GRAIFER, DM
BULYGIN, KN
ZENKOVA, MA
YAMKOVOY, VI
STAHL, J
KARPOVA, GG
机构
[1] RUSSIAN ACAD SCI, NOVOSIBIRSK BIOORGAN CHEM INST, RIBOSOMAL STRUCT & FUNCT LAB, NOVOSIBIRSK 630090, RUSSIA
[2] MAX DELBRUCK CENTRUM MOLEK MED, BERLIN, GERMANY
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 226卷 / 02期
关键词
D O I
10.1111/j.1432-1033.1994.tb20100.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Affinity labeling of human placental 80S ribosomes with mRNA analogs of up to 12 uridyl residues, i.e. alkylating derivatives of oligouridylates bearing either 4-(N-2-chloroethyl-N-methylamino)benzylmethylphosphamide group at the 5'-termini or 2',3'-O-[4-(N-2-chloroethyl-N-methylamino)]benzylidene residue attached to the 3'-termini, in the presence of cognate Phe-tRNA(Phe) has been investigated. All the mRNA analogs modified only the 40S subunit. The fraction of 18S rRNA modified by the mRNA analogs with the alkylating group at the 5'-end decreased dramatically with extension of the reagent oligouridylate moiety. Nucleotides of 18S rRNA alkylated with the mRNA analogs were determined using a reverse transcription technique. For the mRNA analogs with the alkylating groups at the 3'-termini, G1702 and G1763/G1764 were identified as the cross-linking sites. The intensities of the bands corresponding to reverse transcriptase stops depended on the length of the reagent oligouridylate moieties. Cross-linking sites of the mRNA analogs with the alkylating group at the 5'-termini on 18S rRNA were A1023, C1026, C1057 and A1058 for the (pU)(3) and (pU)(4) derivatives and a single nucleotide C1057 for the (pU)(6) one. Ribosomal protein S26 was found as the main target of modification with the same derivatives of (pU)(6) and (pU)(12).
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页码:715 / 723
页数:9
相关论文
共 38 条
[1]   THE MESSENGER-RNA BINDING TRACK IN THE ESCHERICHIA-COLI RIBOSOME FOR MESSENGER-RNAS OF DIFFERENT SEQUENCES [J].
BHANGU, R ;
WOLLENZIEN, P .
BIOCHEMISTRY, 1992, 31 (25) :5937-5944
[2]   A DETAILED MODEL OF THE 3-DIMENSIONAL STRUCTURE OF ESCHERICHIA-COLI 16-S RIBOSOMAL-RNA INSITU IN THE 30-S SUBUNIT [J].
BRIMACOMBE, R ;
ATMADJA, J ;
STIEGE, W ;
SCHULER, D .
JOURNAL OF MOLECULAR BIOLOGY, 1988, 199 (01) :115-136
[3]  
BULYGIN KN, 1992, MOL BIOL+, V26, P556
[4]   RRNA-PROTEIN NEIGHBORHOOD IN ESCHERICHIA-COLI 70-S RIBOSOMES AND 70-S INITIATION COMPLEX - PROBING BY BIFUNCTIONAL PT(II)-CONTAINING REAGENT [J].
CHISTYAKOV, PG ;
ABDUKAYUMOV, MN ;
VENIAMINOVA, AG ;
VLADIMIROV, SN ;
GRAIFER, DM ;
KAZAKOV, SA ;
KARPOVA, GG .
FEBS LETTERS, 1988, 236 (01) :246-250
[5]   G1401 - A KEYSTONE NUCLEOTIDE AT THE DECODING SITE OF ESCHERICHIA-COLI 30S RIBOSOMES [J].
CUNNINGHAM, PR ;
NURSE, K ;
WEITZMANN, CJ ;
NEGRE, D ;
OFENGAND, J .
BIOCHEMISTRY, 1992, 31 (33) :7629-7637
[6]   3 WIDELY SEPARATED POSITIONS IN THE 16S-RNA LIE IN OR CLOSE TO THE RIBOSOMAL DECODING REGION - A SITE-DIRECTED CROSS-LINKING STUDY WITH MESSENGER-RNA ANALOGS [J].
DONTSOVA, O ;
DOKUDOVSKAYA, S ;
KOPYLOV, A ;
BOGDANOV, A ;
RINKEAPPEL, J ;
JUNKE, N ;
BRIMACOMBE, R .
EMBO JOURNAL, 1992, 11 (08) :3105-3116
[7]   ENERGY-TRANSFER EQUILIBRIUM BETWEEN 2 IDENTICAL COPIES OF A RIBOSOME-BOUND FLUORESCENT TRANSFER-RNA ANALOG - IMPLICATIONS FOR THE POSSIBLE STRUCTURE OF CODON-ANTICODON COMPLEXES [J].
FAIRCLOUGH, RH ;
CANTOR, CR .
JOURNAL OF MOLECULAR BIOLOGY, 1979, 132 (04) :587-601
[8]   IDENTIFICATION OF A SITE ON 18-S RIBOSOMAL-RNA OF HUMAN PLACENTA RIBOSOMES IN THE REGION OF THE MESSENGER-RNA BINDING CENTER [J].
GRAIFER, DM ;
ZENKOVA, MA ;
MALYGIN, AA ;
MAMAEV, SV ;
MUNDUS, DA ;
KARPOVA, GG .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 214 (01) :121-128
[9]   INTERACTION OF HUMAN AND ESCHERICHIA-COLI TRANSFER RNAPHE WITH HUMAN 80S RIBOSOMES IN THE PRESENCE OF OLIGOURIDYLATE AND POLYURIDYLATE TEMPLATES [J].
GRAIFER, DM ;
NEKHAI, SY ;
MUNDUS, DA ;
FEDOROVA, OS ;
KARPOVA, GG .
BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1171 (01) :56-64
[10]   FUNCTIONAL TOPOGRAPHY OF HUMAN RIBOSOMES AS STUDIED BY AFFINITY LABELING WITH REACTIVE MESSENGER-RNA ANALOGS [J].
KARPOVA, GG ;
GRAIFER, DM ;
MALYGIN, AA ;
MUNDUS, DA ;
ZENKOVA, MA ;
MAMAEV, SV .
BIOCHIMIE, 1992, 74 (04) :373-380