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
[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).
机构:
ST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USAST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USA
BHANGU, R
;
WOLLENZIEN, P
论文数: 0引用数: 0
h-index: 0
机构:
ST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USAST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USA
机构:
ST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USAST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USA
BHANGU, R
;
WOLLENZIEN, P
论文数: 0引用数: 0
h-index: 0
机构:
ST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USAST LOUIS UNIV, MED CTR, EDWARD A DOISY DEPT BIOCHEM & MOLEC BIOL, ST LOUIS, MO 63104 USA