% post CLS lifesci course on 15th January
% draft. Remaining items are AACon, RNA visualization/editing and Protein disorder analysis exercises.
-8th December 2016
+15th February 2017
\end{center}
building, principal components analysis, physico-chemical property conservation
and sequence consensus analyses are built into the program. Web services enable
Jalview to access online alignment and secondary structure prediction programs,
-as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation. Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem. Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout. Alignment views are dynamically linked with Jmol structure displays, a tree viewer and spatial cluster display, facilitating interactive exploration of the alignment's structure. The application provides its own Jalview project file format in order to store the current state of an alignment and analysis windows. Jalview also provides WYSIWIG\footnote{WYSIWIG: What You See Is What You Get.} style figure generation capabilities for the preparation of alignments for publication.
+as well as to retrieve protein and nucleic acid sequences, alignments, protein structures and sequence annotation.
+Sequences, alignments, trees, structures, features and alignment annotation may also be exchanged with the local filesystem.
+Multiple visualizations of an alignment may be worked on simultaneously, and the user interface provides a comprehensive set of controls for colouring and layout.
+Alignment views are dynamically linked with Jmol and UCSF Chimera\footnote{UCSF Chimera needs to be installed separately. It is available free for academic use from \url{https://www.cgl.ucsf.edu/chimera/download.html}.} structure displays,
+a tree viewer and spatial cluster display, facilitating interactive exploration of the alignment's structure. The application provides its own Jalview project file format in order
+to store the current state of an alignment and analysis windows. Jalview also provides WYSIWIG\footnote{WYSIWIG: What You See Is What You Get.} style
+ figure generation capabilities for the preparation of alignments for publication.
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=5.8in]{images/jvcapabilities.pdf}
\subsection{About this Tutorial }
This tutorial is written in a manual format with short exercises where
-appropriate, typically at the end of each section. This chapter concerns the
+appropriate, typically at the end of each section. The first few sections concerns the
basic operation of Jalview and should be sufficient for those who want to
launch Jalview (Section \ref{startingjv}), open an alignment (Section
\ref{loadingseqs}), perform basic editing (Section
\ref{selectingandediting}), colouring (Section \ref{colours}), and produce
publication and presentation quality graphical output (Section \ref{layoutandoutput}).
-In addition, the manual covers the additional visualization and
-analysis techniques available in Jalview. This includes working
-with the embedded Jmol molecular structure viewer, building and viewing trees and PCA
-plots, and using trees for sequence conservation analysis. An overview of
+The remaining sections of the manual cover the visualization and
+analysis techniques available in Jalview. These include working
+with the embedded Jmol molecular structure viewer (or UCSF Chimera), building
+and viewing trees and Principal Components Analysis (PCA) plots, and using
+trees for sequence conservation analysis. An overview of
the Jalview Desktop's webservices is given in Section \ref{jvwebservices}, and
the alignment and secondary structure prediction services are described
-in detail in Sections \ref{msaservices} and \ref{protsspredservices}. Section \ref{featannot} details the creation and visualization of sequence
-and alignment annotation. Section \ref{workingwithnuc} discusses
+in detail in Sections \ref{msaservices} and \ref{protsspredservices}
+respectively.
+Section \ref{featannot} details the creation and visualization of sequence and alignment annotation. Section \ref{workingwithnuc} discusses
specific features of use when working with nucleic acid sequences, such as translation and linking to protein
coding regions, and the display and analysis of RNA secondary structure.
Keystrokes using the special non-symbol keys are represented in the tutorial by
enclosing the pressed keys with square brackets ({\em e.g.} [RETURN] or [CTRL]).
-Keystroke combinations are combined with a `-' symbol ({\em e.g.} [CTRL]-C means
-press [CTRL] and the `C' key) simultaneously.
+Keystroke combinations are denoted with a `-' symbol ({\em
+e.g.} [CTRL]-C means press [CTRL] and the `C' key simultaneously).
Menu options are given as a path from the menu
that contains them - for example {\sl File $\Rightarrow$ Input Alignment
Jalview site. This behaviour can be switched off in the Jalview Desktop
preferences dialog by unchecking the open file option.
This alignment will look like the one in Figure \ref{startpage} (taken
-from Jalview version 2.7).
+from Jalview version 2.10.1).
%[figure 3 ]
\begin{figure}[htbp]
Archives and mailing list
subscription details can be found in the Jalview web site's \href{http://www.jalview.org/community}{community section}.
+
\section{Navigation}
\label{jvnavigation}
The major features of the Jalview Desktop are illustrated in Figure \ref{anatomy}. The alignment window is the primary window for editing and visualization, and can contain several independent views of the alignment being worked with. The other windows (Trees, Structures, PCA plots, etc) are linked to a specific alignment view. Each area of the alignment window has a separate context menu accessed by clicking the right mouse button.
Jalview has two navigation and editing modes: {\bf normal mode}, where
editing and navigation is performed using the mouse, and {\bf cursor mode}
where editing and navigation are performed using the keyboard. The {\bf F2 key}
- is used to switch between these two modes. With a Mac as the F2 is
- often assigned to screen brightness, one may often need to type {\bf function
- [Fn] key with F2} function
- [Fn]-F2.
+ is used to switch between these two modes.
+
+ {\em Note:} On MacBooks and other laptops with compact keyboards, you may need
+ to press the {\bf function key [Fn]} when pressing any of the numbered function
+ keys. So to toggle between keyboard and normal mode, press [Fn]-[F2].
+
\begin{figure}[htb]
\begin{center}
Rapid movement to specific positions is accomplished as listed below:
\begin{list}{$\circ$}{}
\item {\bf Jump to Sequence {\sl n}:} Type a number {\sl n} then press [S] to
-move to sequence (row). {\sl n}
+move to sequence (row) {\sl n}.
\item {\bf Jump to Column {\sl n}:} Type a number {\sl n} then press [C] to move to column {\sl n} in the alignment.
\item {\bf Jump to Residue {\sl n}:} Type a number {\sl n} then press [P] to move to residue number {\sl n} in the current sequence.
\item {\bf Jump to column {\sl m} row {\sl n}:} Type the column number {\sl m}, a comma, the row number {\sl n} and press [RETURN].
`Paste to new window' option in the menu that appears. The other is to select
{\sl File $\Rightarrow$ Input Alignment $\Rightarrow$ From Textbox} from the
main menu, paste the sequences into the text window that will appear, and select
-{sl New Window} (Figure \ref{loadtext}). In both cases, presuming that they are
+{\sl New Window} (Figure \ref{loadtext}). In both cases, presuming that they are
in the right format, Jalview will happily read them into a new alignment window.
%[fig 8]
also be output to a textbox using the output functions in the pop-up menu
obtained by right clicking the current selection. The textbox enables quick
manual editing of the alignment prior to importing it into a new window (using
-the [New Window] button) or saving to a file with the {\sl File $\Rightarrow$
-Save As } pulldown menu option from the text box.
+the {\sl New Window} button) or saving to a file with the {\sl File
+$\Rightarrow$ Save As } pulldown menu option from the text box.
\section{Reordering an Alignment}
Sequence reordering is simple. Highlight the sequences to be moved then press the up or down arrow keys as appropriate (Figure \ref{reorder}). If you wish to move a sequence up past several other sequences it is often quicker to select the group past which you want to move it and then move the group rather than the individual sequence.
Gaps can be removed by dragging the residue to the immediate right of the gap
leftwards whilst holding down [SHIFT] (for single sequences) or [CTRL] (for a group of sequences).
-\subsection{Sliding Sequences}
-Pressing the [$\leftarrow$] or [$\rightarrow$] arrow keys when one or more
-sequences are selected will ``slide'' the entire selected sequences to the left
-or right (respectively). Slides occur regardless of the region selection -
-which, for example, allows you to easily reposition misaligned subfamilies
-within a larger alignment.
-% % better idea to introduce hiding sequences, and use the invert selection, hide
-% others, to simplify manual alignment construction
-
-\subsection{Editing in Cursor mode}
-Gaps can be easily inserted when in cursor mode (toggled with [F2]) by
-pressing [SPACE]. Gaps will be inserted at the cursor, shifting the residue
-under the cursor to the right. To insert {\sl n} gaps type {\sl n} and then
-press [SPACE]. To insert gaps into all sequences of a group, use [CTRL]-[SPACE]
-or [SHIFT]-[SPACE] (both keys held down together).
-
-Gaps can be removed in cursor mode by pressing [BACKSPACE]. First make sure you
-have everything unselected by pressing ESC. The gap under the cursor will be
-removed. To remove {\sl n} gaps, type {\sl n} and then press [BACKSPACE]. Gaps
-will be deleted up to the number specified. To delete gaps from all sequences of
-a group, press [CTRL]-[BACKSPACE] or [SHIFT]-[BACKSPACE] (both keys held down
-together). Note that the deletion will only occur if the gaps are in the same
-columns in all sequences in the selected group, and those columns are to the
-right of the selected residue.
-
\newpage
\exercise{Editing Alignments}
backwards and replay the edits you have made.}
}
+\subsection{Sliding Sequences}
+Pressing the [$\leftarrow$] or [$\rightarrow$] arrow keys when one or more
+sequences are selected will ``slide'' the entire selected sequences to the left
+or right (respectively). Slides occur regardless of the region selection -
+which, for example, allows you to easily reposition misaligned subfamilies
+within a larger alignment.
+% % better idea to introduce hiding sequences, and use the invert selection, hide
+% others, to simplify manual alignment construction
+
+\subsection{Editing in Cursor mode}
+Gaps can be easily inserted when in cursor mode (toggled with [F2]) by
+pressing [SPACE]. Gaps will be inserted at the cursor, shifting the residue
+under the cursor to the right. To insert {\sl n} gaps type {\sl n} and then
+press [SPACE]. To insert gaps into all sequences of a group, use [CTRL]-[SPACE]
+or [SHIFT]-[SPACE] (both keys held down together).
+
+Gaps can be removed in cursor mode by pressing [BACKSPACE]. First make sure you
+have everything unselected by pressing ESC. The gap under the cursor will be
+removed. To remove {\sl n} gaps, type {\sl n} and then press [BACKSPACE]. Gaps
+will be deleted up to the number specified. To delete gaps from all sequences of
+a group, press [CTRL]-[BACKSPACE] or [SHIFT]-[BACKSPACE] (both keys held down
+together). Note that the deletion will only occur if the gaps are in the same
+columns in all sequences in the selected group, and those columns are to the
+right of the selected residue.
+
\exercise{Keyboard Edits}
{This continues on from the previous exercise, and recreates the final part of the example ferredoxin
} \parbox[c]{3in}{
\includegraphics[width=2.75in]{images/col_rnahelix.pdf} }
+\subsubsection{User Defined}
+This dialog allows the user to create any number of named colour schemes at
+will. Any residue may be assigned any colour. The colour scheme can then be
+named. If you save the colour scheme, this name will appear on the Colour menu
+(Figure \ref{usercol}).
+
+
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=2.5in]{images/col_user1.pdf}
+\includegraphics[width=2in]{images/col_user2.pdf}
+\includegraphics[width=1.75in]{images/col_user3.pdf}
+\caption{{\bf Creation of a user defined colour scheme.} Residue types are assigned colours (left). The profile is saved (center) and can then be accessed {\sl via} the {\sl Colour} menu (right).}
+\label{usercol}
+\end{center}
+\end{figure}
+
\exercise{Colouring Alignments}{
\label{color}
Note: Before you begin this exercise, ensure that the {\sl Apply Colour
{\bf See the video at: \url{http://www.jalview.org/training/Training-Videos}.}
}
-\subsubsection{User Defined}
-This dialog allows the user to create any number of named colour schemes at
-will. Any residue may be assigned any colour. The colour scheme can then be
-named. If you save the colour scheme, this name will appear on the Colour menu
-(Figure \ref{usercol}).
-
-
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=2.5in]{images/col_user1.pdf}
-\includegraphics[width=2in]{images/col_user2.pdf}
-\includegraphics[width=1.75in]{images/col_user3.pdf}
-\caption{{\bf Creation of a user defined colour scheme.} Residue types are assigned colours (left). The profile is saved (center) and can then be accessed {\sl via} the {\sl Colour} menu (right).}
-\label{usercol}
-\end{center}
-\end{figure}
-
\exercise{User Defined Colour Schemes}{
\exstep{Load a sequence alignment. Select the alignment menu option {\sl Colour $\Rightarrow$ User Defined}. A dialog window will open.}
but are shown mapped on to specific residues within the alignment.
Features and annotation can be interactively created, or retrieved from external
-data sources. Webservices like JNet (see \ref{jpred} above) can be used to analyse a
-given sequence or alignment and generate annotation for it.
+data sources. Webservices like JPred (see \ref{jpred} above) can be used to
+analyse a given sequence or alignment and generate annotation for it.
\section{Conservation, Quality and Consensus Annotation}
If a sequence is verified, then the start/end numbering will be adjusted to match the Uniprot record.
\subsubsection{Rate of Feature Retrieval}
-Feature retrieval can take some time if a large number of sources is selected and if the alignment
-contains a large number of sequences.
+Feature retrieval can take some time if a large number of sources are selected
+and if the alignment contains a large number of sequences.
As features are retrieved, they are immediately added to the current alignment view.
The retrieved features are shown on the sequence and can be customised as described previously.
McWilliam H, Remmert M, Soding J, Thompson JD, Higgins DG (2011) {\sl Molecular
Systems Biology} {\bf 7} 539
\href{http://dx.doi.org/10.1038/msb.2011.75}{doi:10.1038/msb.2011.75}} Of these,
-T-COFFEE is slow but the accurate. ClustalW is historically
+T-COFFEE is slow but accurate. ClustalW is historically
the most widely used. Muscle is fast and probably best for
smaller alignments. MAFFT is probably the best for large alignments,
however Clustal Omega, released in 2011, is arguably the fastest and most
On calculating a tree, a new window opens (Figure \ref{trees1}) which contains
the tree. Various display settings can be found in the tree window {\sl View}
-menu, including font, scaling and label display options, and the {\sl File
+menu, including font, scaling and label display options. The {\sl File
$\Rightarrow$ Save As} submenu contains options for image and Newick file
export. Newick format is a standard file format for trees which allows them to
be exported to other programs. Jalview can also read in external trees in
\subsection{Configuring the default structure viewer}
\label{configuring3dviewer}
-To configure which one is used when creating a new
+To configure which viewer is used when creating a new
structure view, open the Structures preferences window {\sl via} {\sl Tools $\Rightarrow$ Preferences\ldots} and
select either JMOL or CHIMERA as the default viewer. If you select Chimera,
Jalview will search for the installed program, and if it cannot be found,
When the structure chooser is first opened, if no database identifiers are
available, Jalview will attempt to discover identifiers for the sequence and from there discover any
associated PDB structures. This can take a few seconds for each sequence and
-while be performed for all selected sequences. After this is done, you can see
-added database references in a tool tip by mousing over the sequence ID.
-\footnote{Tip:
-The sequence ID tooltip can often become large for heavily cross referenced sequence IDs. Use the {\sl View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$ } submenu to disable the display of database cross references or non-positional features. } now shows the Uniprot ID and any associated PDB structures.
+will be performed for all selected sequences. After this is done, you can see
+the added database references in a tool tip by mousing over the sequence
+ID\footnote{Tip:
+If sequence ID tooltip obscures your view, then use the {\sl View $\Rightarrow$ Sequence ID Tooltip $\Rightarrow$ }
+submenu option to disable the display of database cross references or non-positional
+features. }, now shows the Uniprot ID and any associated PDB structures.
%
% \begin{figure}[htbp]
% \begin{center}
in the structure viewer can be saved as an EPS or PNG with the {\sl File
$\Rightarrow$ Save As $\Rightarrow$ \ldots} submenu, which also allows the raw
data to be saved as PDB format. The mapping between the structure and the
-sequence (How well and which parts of the structure relate to the sequence) can
+sequence (how well and which parts of the structure relate to the sequence) can
be viewed with the {\sl File $\Rightarrow$ View Mapping} menu option.
\subsubsection{Using the Jmol Visualization Interface }
views that colour different areas or domains of the alignment. This option is
further explored in Section \ref{complexstructurecolours}.
-\begin{figure}[htbp]
-\begin{center}
-\includegraphics[width=5.5in]{images/mviewstructurecol.pdf}
-\caption{{\bf Choosing a different view for colouring a structure display}
-Browsing the {\sl View $\Rightarrow$ Colour by ..} menu provides full control
-of which alignment view is used to colour structures when the {\sl Colours
-$\Rightarrow$ By Sequence} option is selected.}
-\label{mviewstructurecol}
-\end{center}
-\end{figure}
\exercise{Aligning Structures using the Ferredoxin
Sequence Alignment}{\label{superpositionex}
Which view do you think give the best 3D superposition, and why ?} }
+\begin{figure}[htbp]
+\begin{center}
+\includegraphics[width=5.5in]{images/mviewstructurecol.pdf}
+\caption{{\bf Choosing a different view for colouring a structure display}
+Browsing the {\sl View $\Rightarrow$ Colour by ..} menu provides full control
+of which alignment view is used to colour structures when the {\sl Colours
+$\Rightarrow$ By Sequence} option is selected.}
+\label{mviewstructurecol}
+\end{center}
+\end{figure}
+
\subsubsection{Colouring Complexes}
\label{complexstructurecolours}
The ability to control which multiple alignment view is used to colour
\begin{center}
\includegraphics[width=2.25in]{images/jpred1.pdf}
\includegraphics[width=3in]{images/jpred2.pdf}
-\caption{{\bf Secondary Structure Prediction} Status (left) and results (right) windows for JNet predictions. }
+\caption{{\bf Secondary Structure Prediction} Status (left) and results (right)
+windows for JPred predictions. }
\label{jpred}
\end{center}
\end{figure}
Jpred is launched in the same way as the other web services. Select {\sl Web
-Service $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet
+Service $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JPred
Secondary Structure Prediction}\footnote{JNet is the Neural Network based
secondary structure prediction method that the JPred server uses.} from the
alignment window menu (Figure \ref{jpred}).
predictions are included as annotations. Consult the Jpred documentation for
information on interpreting these results.
-\subsection{Hidden Columns and JNet Predictions}
+\subsection{Hidden Columns and JPred Predictions}
\label{hcoljnet}
Hidden columns can be used to exclude parts of a sequence or profile from the
input sent to the JNet service. For instance, if a sequence is known to include
reording and hiding autocalculated and sequence associated annotation. }
\exstep{ Open the alignment at \url{http://www.jalview.org/tutorial/alignment.fa}. Select the sequence {\sl FER\_MESCR} by
-clicking on the sequence ID. Then select {\sl Web Service $\Rightarrow$ Secondary Structure Prediction $\Rightarrow$ JNet Secondary Structure Prediction}
-from the alignment window menu. A status window will appear and after some time (about 2-4 min) a new window with the JPred prediction will appear.
+clicking on the sequence ID. Then select {\sl Web Service $\Rightarrow$
+Secondary Structure Prediction $\Rightarrow$ JPred Secondary Structure
+Prediction} from the alignment window menu. A status window will appear and after some time (about 2-4 min) a new window with the JPred prediction will appear.
Note that the number of sequences in the results window is many more than in the original alignment as
-JNet performs a PSI-BLAST search to expand the prediction dataset. The results
-from the prediction are visible in the annotation panel. Jnet secondary
+JPred performs a PSI-BLAST search to expand the prediction dataset. The results
+from the prediction are visible in the annotation panel. JPred secondary
structure prediction annotations are examples of sequence-associated alignment annotation. }
% TODO: check how long this takes - about 2 mins once it gets on the cluster.
\exstep{
-Select a different sequence and perform a JNet prediction in the same way. There will probably be minor differences in the predictions.
+Select a different sequence and perform a JPred prediction in the same way.
+There will probably be minor differences in the predictions.
}
\exstep{
Select the sequence used in the second sequence prediction by clicking on its
by right clicking the mouse to open the context menu, and select the {\sl Add
Reference Annotation} option.
-{\bf All} the JNet predictions for the sequences will now be visible in the
+{\bf All} the JPred predictions for the sequences will now be visible in the
original alignment window.}
{\bf Homework:} Go back to the last step of exercise \ref{annotatingalignex} and
follow the instructions to view the Jalview annotations file created from the annotations
-generated by the JPred server for your sequence.
-
-}
-
+generated by the JPred server for your sequence.}
\section{Protein Disorder Prediction}
\label{protdisorderpred}
allows access to protein disorder prediction services provided by the configured
JABAWS servers.
-
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=5in]{images/disorderpredannot.pdf}
\end{center}
\end{figure}
-
\subsection{Disorder Prediction Results}
Each service operates on sequences in the alignment to identify regions likely
to be unstructured or flexible, or alternately, fold to form globular domains.
thresholding (described in Section \ref{featureschemes}) can be used to
highlight differences in disorder prediction across aligned sequences.
-
\begin{figure}[htbp]
\begin{center}
\includegraphics[width=5in]{images/disorderpred.pdf}
the associated sequence in the alignment display, and double clicking will
select that sequence.
-
\subsection{Disorder Predictors provided by JABAWS 2.0}
For full details of each predictor and the results that Jalview can display,
please consult
available temperature factors to the alignment {\sl via} the {\sl Sequence ID
Popup $\Rightarrow$ Selection $\Rightarrow$ Add reference annotation} option.}
-\exstep{Apply the IUPred disorder prediction method.}
-\exstep{Use the {\sl Per
+\exstep{Apply the IUPred disorder prediction method. Use the {\sl Per
sequence option} in the {\sl Colour $\Rightarrow$ By annotation \ldots} dialog to shade
-the sequences by the long and short disorder predictors.
-
-{\sl Note how well the regions predicted to be disordered by the methods agree
-with the structure.}
-}
-
-}
+the sequences by the long and short disorder predictors. {\sl Note how well the disordered regions predicted by each method agree
+with the structure.}}}
\chapter{DNA and RNA Sequences}
\label{dnarna}
\subsection{Running your own JABA Server}
You can download and run JABA on your own machine using the `VMWare' or
-VirtualBox virtual machine environments. If you would like to learn how to do
+VirtualBox virtual machine environments. If you would like to do
this, there are full instructions at the
\href{http://www.compbio.dundee.ac.uk/jabaws/}{JABA web site}.
WARNING: This is large (about 300MB) and will take some time to download.
}
\exstep{Unpack the archive's contents to a place on your machine with at least
-2GB of free space.
-
-(On Windows, right click on the archive, and use the 'Extract archive..' option).
+2GB of free space (On Windows, right click on the archive, and use the 'Extract
+archive..' option).
}
\exstep{Open the newly extracted directory and double click the VMWare virtual
machine configuration file (jabaws.vcf). This will launch the VMWare player.
git://source.jalview.org / jalview-manual.git / blobdiff