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NetNES - 1.1

Leucine-rich nuclear export signals (NES) in eukaryotic proteins

The NetNES 1.1 server predicts leucine-rich nuclear export signals (NES) in eukaryotic proteins using a combination of neural networks and hidden Markov models.

Out of Commission

This service is unfortunately out of commission at this time.
The software may still be available for download for academic institutions at the Downloads tab.

Instructions


1. Specify the input sequences

All the input sequences must be in one-letter amino acid code. The allowed alphabet (not case sensitive) is as follows:

A C D E F G H I K L M N P Q R S T V W Y

All the other symbols will be converted to A before processing. The sequences can be input in the following two ways:

  • Paste a single sequence (just the amino acids) or a number of sequences in FASTA format into the upper window of the main server page.

  • Select a FASTA file on your local disk, either by typing the file name into the lower window or by browsing the disk.

Both ways can be employed at the same time: all the specified sequences will be processed. However, there may be not more than 2,000 sequences and 200,000 amino acids in one submission. The sequences should not be shorter than 15 or longer than 7,000 amino acids.


2. Customize your run

The button labelled "Generate graphics" controls whether a graphical output of the prediction is created. By default it is on - click to turn off.


3. Submit the job

Click on the "Submit" button. The status of your job (either 'queued' or 'running') will be displayed and constantly updated until it terminates and the server output appears in the browser window.

At any time during the wait you may enter your e-mail address and simply leave the window. Your job will continue; you will be notified by e-mail when it has terminated. The e-mail message will contain the URL under which the results are stored; they will remain on the server for 24 hours for you to collect them.

Output format


DESCRIPTION

Providing 'Generate graphics' is enabled, server output consists of two parts; one is a listing of all residues with the indiviual scores noted (see column heading), the other is a graphical plot of the values given in the first table. If 'Generate graphics' is deselected the plot will be omitted.

As described in the paper, the prediction server calculates the NES score from the HMM and Artificial Neural Network (ANN) scores but all three values are given for each residue. This allows for manual inspection of possible mispredictions. A wrong prediction is sometimes obtained because either the HMM or the ANN fails to recognize the NES motif completely. In that case an experienced user might be able to spot that from the individual scores.

Even though the ANN is trained to specifically recognize the last (most C-terminal) hydrophobic position of the NES motif, it is often seen that the other hydrophobic positions give rise to peaks in the ANN score. Again, the experienced user might benefit from the additional information in the ANN score and be able to rescue a misprediction.

In summary, if the calculated score 'NES score' exceeds the threshold, then that particular residue is expected to participate in a nuclear export signal. This is denoted with a 'Yes' in the column 'Predicted'.

Note that non-standard amino acid abbreviations (BJOUXZ) are replaced with alanines.

For further information, please refer to the publication or contact us.


EXAMPLE OUTPUT





>REV_HV1C4 - netNES 1.1 prediction
#Seq-Pos-Residue         ANN     HMM     NES   Predicted
#---------------------------------------------------------
REV_HV1C4-1-M           0.121   0.000   0.000      -
REV_HV1C4-2-A           0.081   0.000   0.000      -
REV_HV1C4-3-G           0.109   0.000   0.000      -
REV_HV1C4-4-R           0.089   0.000   0.000      -
REV_HV1C4-5-S           0.097   0.000   0.000      -
REV_HV1C4-6-G           0.096   0.000   0.000      -
REV_HV1C4-7-D           0.090   0.000   0.000      -
REV_HV1C4-8-S           0.084   0.000   0.000      -
REV_HV1C4-9-D           0.087   0.000   0.000      -
REV_HV1C4-10-E          0.125   0.000   0.000      -
REV_HV1C4-11-E          0.078   0.000   0.000      -
REV_HV1C4-12-L          0.168   0.000   0.000      -
REV_HV1C4-13-I          0.086   0.000   0.000      -
REV_HV1C4-14-K          0.096   0.000   0.000      -
REV_HV1C4-15-T          0.098   0.000   0.000      -
REV_HV1C4-16-V          0.094   0.000   0.000      -
REV_HV1C4-17-R          0.114   0.000   0.000      -
REV_HV1C4-18-I          0.193   0.000   0.000      -
REV_HV1C4-19-I          0.101   0.000   0.000      -
REV_HV1C4-20-K          0.081   0.000   0.000      -
REV_HV1C4-21-H          0.236   0.000   0.000      -
REV_HV1C4-22-L          0.445   0.000   0.234      -
REV_HV1C4-23-Y          0.094   0.000   0.000      -
REV_HV1C4-24-Q          0.097   0.000   0.000      -
REV_HV1C4-25-S          0.071   0.000   0.000      -
REV_HV1C4-26-N          0.069   0.000   0.000      -
REV_HV1C4-27-P          0.093   0.000   0.000      -
REV_HV1C4-28-P          0.087   0.000   0.000      -
REV_HV1C4-29-P          0.070   0.000   0.000      -
REV_HV1C4-30-K          0.064   0.000   0.000      -
REV_HV1C4-31-P          0.070   0.000   0.000      -
REV_HV1C4-32-E          0.067   0.000   0.000      -
REV_HV1C4-33-G          0.066   0.000   0.000      -
REV_HV1C4-34-T          0.069   0.000   0.000      -
REV_HV1C4-35-R          0.076   0.000   0.000      -
REV_HV1C4-36-Q          0.082   0.000   0.000      -
REV_HV1C4-37-A          0.087   0.000   0.000      -
REV_HV1C4-38-R          0.074   0.000   0.000      -
REV_HV1C4-39-R          0.070   0.000   0.000      -
REV_HV1C4-40-N          0.075   0.000   0.000      -
REV_HV1C4-41-R          0.077   0.000   0.000      -
REV_HV1C4-42-R          0.077   0.000   0.000      -
REV_HV1C4-43-R          0.081   0.000   0.000      -
REV_HV1C4-44-R          0.087   0.000   0.000      -
REV_HV1C4-45-W          0.087   0.000   0.000      -
REV_HV1C4-46-R          0.075   0.000   0.000      -
REV_HV1C4-47-E          0.086   0.000   0.000      -
REV_HV1C4-48-R          0.087   0.000   0.000      -
REV_HV1C4-49-Q          0.072   0.000   0.000      -
REV_HV1C4-50-R          0.079   0.000   0.000      -
REV_HV1C4-51-R          0.072   0.000   0.000      -
REV_HV1C4-52-I          0.101   0.000   0.000      -
REV_HV1C4-53-H          0.075   0.000   0.000      -
REV_HV1C4-54-S          0.077   0.000   0.000      -
REV_HV1C4-55-I          0.086   0.000   0.000      -
REV_HV1C4-56-S          0.075   0.000   0.000      -
REV_HV1C4-57-A          0.081   0.000   0.000      -
REV_HV1C4-58-W          0.075   0.000   0.000      -
REV_HV1C4-59-L          0.098   0.001   0.000      -
REV_HV1C4-60-L          0.138   0.003   0.000      -
REV_HV1C4-61-S          0.080   0.003   0.000      -
REV_HV1C4-62-T          0.084   0.003   0.000      -
REV_HV1C4-63-C          0.136   0.003   0.000      -
REV_HV1C4-64-L          0.301   0.006   0.000      -
REV_HV1C4-65-G          0.105   0.006   0.000      -
REV_HV1C4-66-R          0.100   0.006   0.000      -
REV_HV1C4-67-S          0.078   0.006   0.000      -
REV_HV1C4-68-A          0.081   0.006   0.000      -
REV_HV1C4-69-E          0.145   0.006   0.000      -
REV_HV1C4-70-P          0.086   0.006   0.000      -
REV_HV1C4-71-V          0.074   0.022   0.000      -
REV_HV1C4-72-P          0.072   0.022   0.000      -
REV_HV1C4-73-L          0.092   0.381   0.091      -
REV_HV1C4-74-Q          0.086   0.380   0.091      -
REV_HV1C4-75-L          0.110   0.998   0.702      Yes
REV_HV1C4-76-P          0.070   0.996   0.696      Yes
REV_HV1C4-77-P          0.082   0.996   0.698      Yes
REV_HV1C4-78-L          0.108   0.996   0.705      Yes
REV_HV1C4-79-E          0.073   0.995   0.701      Yes
REV_HV1C4-80-R          0.234   0.995   0.731      Yes
REV_HV1C4-81-L          0.179   0.995   0.745      Yes
REV_HV1C4-82-T          0.113   0.977   0.735      Yes
REV_HV1C4-83-L          0.872   0.977   1.310      Yes
REV_HV1C4-84-D          0.081   0.000   0.000      -
REV_HV1C4-85-C          0.292   0.000   0.000      -
REV_HV1C4-86-S          0.091   0.000   0.000      -
REV_HV1C4-87-E          0.106   0.000   0.000      -
REV_HV1C4-88-D          0.169   0.000   0.000      -
REV_HV1C4-89-C          0.103   0.000   0.000      -
REV_HV1C4-90-G          0.096   0.000   0.000      -
REV_HV1C4-91-T          0.090   0.000   0.000      -
REV_HV1C4-92-S          0.105   0.000   0.000      -
REV_HV1C4-93-G          0.088   0.000   0.000      -
REV_HV1C4-94-T          0.079   0.000   0.000      -
REV_HV1C4-95-Q          0.077   0.000   0.000      -
REV_HV1C4-96-G          0.073   0.000   0.000      -
REV_HV1C4-97-V          0.069   0.000   0.000      -
REV_HV1C4-98-G          0.075   0.000   0.000      -
REV_HV1C4-99-S          0.087   0.000   0.000      -
REV_HV1C4-100-P         0.072   0.000   0.000      -
REV_HV1C4-101-Q         0.068   0.000   0.000      -
REV_HV1C4-102-I         0.116   0.000   0.000      -
REV_HV1C4-103-L         0.078   0.000   0.000      -
REV_HV1C4-104-V         0.076   0.000   0.000      -
REV_HV1C4-105-E         0.087   0.000   0.000      -
REV_HV1C4-106-S         0.076   0.000   0.000      -
REV_HV1C4-107-P         0.080   0.000   0.000      -
REV_HV1C4-108-A         0.077   0.000   0.000      -
REV_HV1C4-109-V         0.108   0.000   0.000      -
REV_HV1C4-110-L         0.085   0.000   0.000      -
REV_HV1C4-111-E         0.079   0.000   0.000      -
REV_HV1C4-112-S         0.120   0.000   0.000      -
REV_HV1C4-113-G         0.071   0.000   0.000      -
REV_HV1C4-114-T         0.072   0.000   0.000      -
REV_HV1C4-115-K         0.097   0.000   0.000      -
REV_HV1C4-116-E         0.077   0.000   0.000      -
//

References


Analysis and prediction of leucine-rich nuclear export signals
Tanja la Cour, Lars Kiemer, Anne Mølgaard, Ramneek Gupta, Karen Skriver and Søren Brunak
Protein Eng. Des. Sel., 17(6):527-36, 2004.

Abstract

We present a thorough analysis of nuclear export signals and a prediction server, which we have made publicly available. The machine learning prediction method is a significant improvement over the generally used consensus patterns. Nuclear export signals (NESs) are extremely important regulators of the subcellular location of proteins. This regulation has an impact on transcription and other nuclear processes, which are fundamental to the viability of the cell. NESs are studied in relation to cancer, the cell cycle, cell differentiation and other important aspects of molecular biology. Our conclusion from this analysis is that the most important properties of NESs are accessibility and flexibility allowing relevant proteins to interact with the signal. Furthermore, we show that not only the known hydrophobic residues are important in defining a nuclear export signals. We employ both neural networks and hidden Markov models in the prediction algorithm and verify the method on the most recently discovered NESs.

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