- EMDB-3300: Electron cryo-microscopy of a human pre-ribosomal (pre-40S) particle -
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基本情報
登録情報
データベース: EMDB / ID: EMD-3300
タイトル
Electron cryo-microscopy of a human pre-ribosomal (pre-40S) particle
マップデータ
Reconstruction of human pre-40S particle purified by using a HASt-tagged version of the ribosome biogenesis factor LTV1 as bait. the map has been filtered and masked using the RELION1.3 postprocess option.
試料
試料: pre-40S particles purified from human cells by using the tagged ribosome biogenesis factor HASt_LTV1 as bait.
複合体: cytoplasmic precursor of the human small ribosomal subunit
ジャーナル: Nucleic Acids Res / 年: 2016 タイトル: Structure of a human pre-40S particle points to a role for RACK1 in the final steps of 18S rRNA processing. 著者: Natacha Larburu / Christian Montellese / Marie-Françoise O'Donohue / Ulrike Kutay / Pierre-Emmanuel Gleizes / Célia Plisson-Chastang / 要旨: Synthesis of ribosomal subunits in eukaryotes is a complex and tightly regulated process that has been mostly characterized in yeast. The discovery of a growing number of diseases linked to defects ...Synthesis of ribosomal subunits in eukaryotes is a complex and tightly regulated process that has been mostly characterized in yeast. The discovery of a growing number of diseases linked to defects in ribosome biogenesis calls for a deeper understanding of these mechanisms and of the specificities of human ribosome maturation. We present the 19 Å resolution cryo-EM reconstruction of a cytoplasmic precursor to the human small ribosomal subunit, purified by using the tagged ribosome biogenesis factor LTV1 as bait. Compared to yeast pre-40S particles, this first three-dimensional structure of a human 40S subunit precursor shows noticeable differences with respect to the position of ribosome biogenesis factors and uncovers the early deposition of the ribosomal protein RACK1 during subunit maturation. Consistently, RACK1 is required for efficient processing of the 18S rRNA 3'-end, which might be related to its role in translation initiation. This first structural analysis of a human pre-ribosomal particle sets the grounds for high-resolution studies of conformational transitions accompanying ribosomal subunit maturation.
ダウンロード / ファイル: emd_3300.map.gz / 形式: CCP4 / 大きさ: 26.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Reconstruction of human pre-40S particle purified by using a HASt-tagged version of the ribosome biogenesis factor LTV1 as bait. the map has been filtered and masked using the RELION1.3 postprocess option.
ボクセルのサイズ
X=Y=Z: 2.27 Å
密度
表面レベル
登録者による: 0.021 / ムービー #1: 0.021
最小 - 最大
-0.04256002 - 0.22263312
平均 (標準偏差)
0.00185307 (±0.01161947)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
192
192
192
Spacing
192
192
192
セル
A=B=C: 435.84 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
2.27
2.27
2.27
M x/y/z
192
192
192
origin x/y/z
0.000
0.000
0.000
length x/y/z
435.840
435.840
435.840
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-147
-147
-146
NX/NY/NZ
294
294
294
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
192
192
192
D min/max/mean
-0.043
0.223
0.002
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添付データ
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試料の構成要素
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全体 : pre-40S particles purified from human cells by using the tagged r...
全体
名称: pre-40S particles purified from human cells by using the tagged ribosome biogenesis factor HASt_LTV1 as bait.
要素
試料: pre-40S particles purified from human cells by using the tagged ribosome biogenesis factor HASt_LTV1 as bait.
複合体: cytoplasmic precursor of the human small ribosomal subunit
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超分子 #1000: pre-40S particles purified from human cells by using the tagged r...
超分子
名称: pre-40S particles purified from human cells by using the tagged ribosome biogenesis factor HASt_LTV1 as bait. タイプ: sample / ID: 1000 / 集合状態: monomer / Number unique components: 1
分子量
理論値: 1.5 MDa
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超分子 #1: cytoplasmic precursor of the human small ribosomal subunit
超分子
名称: cytoplasmic precursor of the human small ribosomal subunit タイプ: complex / ID: 1 / Name.synonym: pre-40S particle / 組換発現: No / Ribosome-details: ribosome-eukaryote: SSU 40S
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: Human / 組織: Embryonic Kidney / 細胞: HEK 293
分子量
理論値: 1.5 MDa
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
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試料調製
濃度
0.02 mg/mL
緩衝液
pH: 7.6 / 詳細: 10 mM Tris-HCl, 100 mM KCl, 2 mM MgCl2
グリッド
詳細: Glow discharged Quantifoil R2/1 on Cu 300 Mesh grids
凍結
凍結剤: ETHANE / チャンバー内湿度: 95 % / チャンバー内温度: 293 K / 装置: LEICA EM GP / 手法: 1.8 - 2.1'' blot before plunging
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
温度
平均: 100 K
日付
2014年1月23日
撮影
カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 実像数: 3812 / 平均電子線量: 30 e/Å2 詳細: Every image has been acquired in 1s by the direct electron detector (no movies recorded)