1 | #Please insert up references in the next lines (line starts with keyword UP) |
---|
2 | UP arb.hlp |
---|
3 | UP glossary.hlp |
---|
4 | UP rna3d_general.hlp |
---|
5 | |
---|
6 | #Please insert subtopic references (line starts with keyword SUB) |
---|
7 | #SUB subtopic.hlp |
---|
8 | |
---|
9 | # Hypertext links in helptext can be added like this: LINK{ref.hlp|http://add|bla@domain} |
---|
10 | |
---|
11 | #************* Title of helpfile !! and start of real helpfile ******** |
---|
12 | TITLE Superimposing rRNA sequence data, SAI and probes |
---|
13 | |
---|
14 | OCCURRENCE In ARB primary structure editor (ARB_EDIT4) -> RNA3D |
---|
15 | |
---|
16 | DESCRIPTION |
---|
17 | |
---|
18 | SUPERIMPOSING rRNA SEQUENCE DATA |
---|
19 | |
---|
20 | Any rRNA sequence contained in the multiple sequence alignments of ARB primary |
---|
21 | structure editor can be overlaid onto the structure of E. coli in the RNA3D |
---|
22 | window. Desired rRNA sequence to be mapped onto the structure can be selected by |
---|
23 | the left mouse button in the multiple sequence alignment and the selected rRNA |
---|
24 | sequence will be instantly mapped onto the master structure. |
---|
25 | |
---|
26 | The selected rRNA sequence is annotated with mutation (base substitutions), |
---|
27 | insertion and deletion information at each site as compared to the master |
---|
28 | sequence (E. coli). |
---|
29 | |
---|
30 | For the regions where the sequences are aligned without deletion or insertion, |
---|
31 | direct base substitution (mutation) is applied. Because the Câ---Câ distance |
---|
32 | is essentially the same (~10.2 Ã
) in all Watson-Crick base pairs (Watson and |
---|
33 | Crick, 1953), this simple procedure preserves the base pairing and the double |
---|
34 | helical structure while substituting the bases. Although there do exist the |
---|
35 | requirement of structural adjustments for non-Watson-Crick base pairs, currently, |
---|
36 | simple base substitutions are kept because the development of new models to |
---|
37 | achieve the necessary structural adjustments is out of the scope of the RNA3D |
---|
38 | tool. |
---|
39 | |
---|
40 | In the regions where the alignment (of selected rRNA sequence) involves |
---|
41 | insertions, the respective insertion points to corresponding E. coli base |
---|
42 | position in the alignment are shown as down arrows in the crystal structure. The |
---|
43 | number of insertions and the participating nucleotides can also be displayed at |
---|
44 | the insertion points. |
---|
45 | |
---|
46 | In the case of regions, where deletions are observed in the alignment |
---|
47 | corresponding to the master sequence (E. coli), respective sites in the crystal |
---|
48 | structure are indicated as deleted. |
---|
49 | |
---|
50 | DISPLAY OPTIONS |
---|
51 | |
---|
52 | ENABLE MAPPING: |
---|
53 | |
---|
54 | Checking this box will enable the mapping or overlaying of any information |
---|
55 | onto the molecule globally. It is very useful to swiftly switching off |
---|
56 | mapping information. |
---|
57 | |
---|
58 | MAP SELECTED SPECIES: |
---|
59 | |
---|
60 | This check box will enable mapping rRNA sequence data contained in the |
---|
61 | multiple alignments onto the 3D molecule. |
---|
62 | |
---|
63 | DISPLAY BASE DIFFERENCE: |
---|
64 | |
---|
65 | Enabling this check box will display the substitutions or mutations |
---|
66 | observed with respect to E.coli sequence onto 16S rRNA 3D structure. |
---|
67 | |
---|
68 | DISPLAY BASE POSITION: |
---|
69 | |
---|
70 | Base positions corresponding to the observed substitutions or mutations in |
---|
71 | the mapped rRNA sequence are displayed by enabling this check box. |
---|
72 | |
---|
73 | DISPLAY DELETIONS: |
---|
74 | |
---|
75 | Enabling this check box will display deletions in mapped rRNA sequence |
---|
76 | with respect to E.coli reference sequence data. |
---|
77 | |
---|
78 | DISPLAY INSERTIONS: |
---|
79 | |
---|
80 | Enabling this check box will display insertions in mapped rRNA sequence |
---|
81 | with respect to E.coli reference sequence data. By checking 'Bases' box, |
---|
82 | the number of insertions along with the actual bases or residues is |
---|
83 | displayed at the insertion points. |
---|
84 | |
---|
85 | DISPLAY MISSING BASES: |
---|
86 | |
---|
87 | Bases or residues which are presumed to be missing in the rRNA sequence |
---|
88 | alignments when comparing with the consensus model and/or during manual |
---|
89 | curation, can be visualized in the 3D structure. Missing bases denoted as |
---|
90 | dots ('.') in the multiple sequence alignments are mapped onto the rRNA 3D |
---|
91 | structure as question marks ('?') by enabling this check box. Such missing |
---|
92 | bases are more often attributed to errors during sequencing. |
---|
93 | |
---|
94 | Color settings related to mapped sequence data including insertions, deletions, |
---|
95 | mutations, and missing residues can be changed using 'Color Settings' of the main |
---|
96 | RNA3D window. |
---|
97 | |
---|
98 | MAPPING OLIGO-NUCLEOTIDE PROBES: |
---|
99 | |
---|
100 | The localization of the proposed oligo-nucleotide probe targets can be |
---|
101 | visualized in customizable background colors with in the rRNA crystal |
---|
102 | structure. Using the navigation capabilities of RNA3D tool (see |
---|
103 | âNavigationâ section), one can get an idea about the probable binding |
---|
104 | site of the proposed probe with respect to the structural conformation of |
---|
105 | rRNA. |
---|
106 | |
---|
107 | Oligo-nucleotide probes are designed using integrated Probe Design and |
---|
108 | Probe Match tools of ARB. The selected oligo-nucleotide probe in probe |
---|
109 | match window is directly mapped onto the rRNA 3D structure by enabling |
---|
110 | âMap Search Patternsâ check box. |
---|
111 | |
---|
112 | OVERLAYING SEQUENCE ASSOCIATED INFORMATION (SAI): |
---|
113 | |
---|
114 | Various column statistics like sequence consensus, base frequency, |
---|
115 | positional variability based on parsimony method and any other user |
---|
116 | defined column statistics that are performed on the sequence alignments |
---|
117 | can be readily overlaid onto the 3D structure. |
---|
118 | |
---|
119 | Once the column statistics are performed, the user can define the color |
---|
120 | translation table for the chosen SAI in the ARB primary structure editor |
---|
121 | (see âView | Visualize SAIsâ menu). Different colors (up to 10 colors) |
---|
122 | can be set to the values or characters stored in the SAI to visualize in |
---|
123 | the molecular structure. The molecule can be re-colored using new settings |
---|
124 | anytime by clicking the color palate button (using Color Settings in RNA3D |
---|
125 | window). |
---|
126 | |
---|
127 | By enabling the âMap Sequence Associated Informationâ check box, the |
---|
128 | transformed data is readily overlaid onto the rRNA 3D structure. Any |
---|
129 | change in the SAIs and respective color transformations can be reapplied |
---|
130 | by clicking ârefreshâ button. |
---|
131 | |
---|
132 | NOTES None |
---|
133 | |
---|
134 | EXAMPLES None |
---|
135 | |
---|
136 | WARNINGS None |
---|
137 | |
---|
138 | BUGS No bugs known |
---|