ジャーナル: J Virol / 年: 2011 タイトル: Residues of the UL25 protein of herpes simplex virus that are required for its stable interaction with capsids. 著者: Shelley K Cockrell / Jamie B Huffman / Katerina Toropova / James F Conway / Fred L Homa / 要旨: The herpes simplex virus 1 (HSV-1) UL25 gene product is a minor capsid component that is required for encapsidation, but not cleavage, of replicated viral DNA. UL25 is located on the capsid surface ...The herpes simplex virus 1 (HSV-1) UL25 gene product is a minor capsid component that is required for encapsidation, but not cleavage, of replicated viral DNA. UL25 is located on the capsid surface in a proposed heterodimer with UL17, where five copies of the heterodimer are found at each of the capsid vertices. Previously, we demonstrated that amino acids 1 to 50 of UL25 are essential for its stable interaction with capsids. To further define the UL25 capsid binding domain, we generated recombinant viruses with either small truncations or amino acid substitutions in the UL25 N terminus. Studies of these mutants demonstrated that there are two important regions within the capsid binding domain. The first 27 amino acids are essential for capsid binding of UL25, while residues 26 to 39, which are highly conserved in the UL25 homologues of other alphaherpesviruses, were found to be critical for stable capsid binding. Cryo-electron microscopy reconstructions of capsids containing either a small tag on the N terminus of UL25 or the green fluorescent protein (GFP) fused between amino acids 50 and 51 of UL25 demonstrate that residues 1 to 27 of UL25 contact the hexon adjacent to the penton. A second region, most likely centered on amino acids 26 to 39, contacts the triplex that is one removed from the penton. Importantly, both of these UL25 capsid binding regions are essential for the stable packaging of full-length viral genomes.
ダウンロード / ファイル: emd_1905.map.gz / 形式: CCP4 / 大きさ: 111 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Surface view of HSV-1 C-capsid where the UL25 protein has been genetically tagged with a tandem affinity purification (TAP) tag at its N-terminus.
ボクセルのサイズ
X
Y
Z
EMDB情報
2.54
2.54
2.54
CCP4マップ ヘッダ情報
2.54
2.54
2.54
EM Navigator ムービー #1
5.08
5.08
5.08
密度
表面レベル
登録者による: 0.8 / ムービー #1: 0.8
最小 - 最大
-7.12019396 - 6.77211857
平均 (標準偏差)
0.00000001 (±1.0)
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
310
310
310
Spacing
310
310
310
セル
A=B=C: 787.39996 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
2.54
2.54
2.54
M x/y/z
310
310
310
origin x/y/z
0.000
0.000
0.000
length x/y/z
787.400
787.400
787.400
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-56
-56
-55
NX/NY/NZ
112
112
112
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
310
310
310
D min/max/mean
-7.120
6.772
0.000
-
添付データ
-
試料の構成要素
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全体 : HSV-1 C-capsid with the UL25 protein labeled with a tandem affini...
全体
名称: HSV-1 C-capsid with the UL25 protein labeled with a tandem affinity purification (TAP) tag at amino acid 50.
要素
試料: HSV-1 C-capsid with the UL25 protein labeled with a tandem affinity purification (TAP) tag at amino acid 50.
ウイルス: Human herpesvirus 1 strain KOS (ヘルペスウイルス)
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超分子 #1000: HSV-1 C-capsid with the UL25 protein labeled with a tandem affini...
超分子
名称: HSV-1 C-capsid with the UL25 protein labeled with a tandem affinity purification (TAP) tag at amino acid 50. タイプ: sample / ID: 1000 / Number unique components: 1
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超分子 #1: Human herpesvirus 1 strain KOS
超分子
名称: Human herpesvirus 1 strain KOS / タイプ: virus / ID: 1 / Name.synonym: HSV-1 / NCBI-ID: 10306 / 生物種: Human herpesvirus 1 strain KOS / ウイルスタイプ: VIRION / ウイルス・単離状態: STRAIN / ウイルス・エンベロープ: No / ウイルス・中空状態: No / Syn species name: HSV-1
宿主
生物種: Homo sapiens (ヒト) / 別称: VERTEBRATES
ウイルス殻
Shell ID: 1 / 直径: 1250 Å / T番号(三角分割数): 16
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実験情報
-
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
-
試料調製
緩衝液
pH: 7.5 / 詳細: 500 mM NaCl, 10 mM Tris, 1 mM EDTA
凍結
凍結剤: ETHANE / チャンバー内湿度: 85 % / 装置: FEI VITROBOT MARK III / 詳細: Vitrification instrument: Vitrobot mark III / 手法: 6s blot prior to plunging