[Binding of human ribosomal protein S5 with the 18S rRNA fragment 1203-1236/1521-1698].
Human ribosomal protein S5 is a homologue of prokaryotic ribosomal protein S7 that binds to the 16S rRNA region formed by helices H28-30 and H41-43 and contacts this site within the 30S ribosomal subunit. To date, one of the most available approaches to study the structure of the eukaryotic ribosome is based on studying binding of rRNA transcripts with recombinant ribosomal proteins. In the present work, we report studying interaction of human recombinant protein S5 with an RNA transcript "corresponding to the region 1203-1236/1521-1698 (helices H28-30 and H41-43) of human 18S rRNA. The homologous region of 16S rRNA is known to bind protein S7. The apparent association constant of S5 with the transcript was determined. Nucleotide residues of the transcript changing their accessibility for cleavage with RNases T1 and T2 in the presence of S5 were determined by the enzymatic footprinting. It was shown that these nucleotides are located preferentially in the internal loop of the fragment formed by basal parts of the helices H29, H30 and H41 that agrees with the data on the S7 binding to 16S rRNA and on its location within the 30S ribosomal subunit. Besides, we also found nucleotides in the transcript regions corresponding to 16S rRNA regions that do not contact S7 in the 30S subunit.
Ian'shina
D D
DD
Malygin
A A
AA
Karpova
G G
GG
D006801:Humans; D008958:Models, Molecular; D011485:Protein Binding; D023302:Protein Footprinting; D017434:Protein Structure, Tertiary; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000378:metabolism; D012337:RNA, Ribosomal, 18S / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000378:metabolism
D012336:RNA, Ribosomal, 16S; D012337:RNA, Ribosomal, 18S; D011994:Recombinant Proteins; D012269:Ribosomal Proteins; C030831:ribosomal protein S5