Question

AnnotationForge::makeOrgPackage GO Ids mistake

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Saber_J • 0

@e5bd6104

Last seen 7 weeks ago

China

Hello everyone, I'm trying to create a custom annotation for Locusta migratoria and then run an enrichment analysis with clusterProfiler. However, I met an error when running AnnotationForge::makeorgPackage(), which returns the following error:

Populating genes table:
genes table filled
Populating gene_info table:
gene_info table filled
Populating go table:
go table filled
table metadata filled
Error in makeOrgDbFromDataFrames(data, tax_id, genus, species, dbFileName,  : 
  'goTable' GO Ids must be formatted like 'GO:XXXXXXX'

My code is :

AnnotationForge::makeOrgPackage(gene_info=gene_info,
                                go=gene2go,
                                maintainer='liXY <XYZ_zz@163.com>',
                                author='liXY',
                                outputDir="E:/reKEGG/LocalOrgdb/20220416",
                                tax_id=7004,
                                genus='Locusta',
                                species='migratoria',
                                goTable="go",
                                version="1.0")

2 files I used:

> head(gene2go)
         GID         GO EVIDENCE
1 LOCMI17615 GO:0000003      IEA
2 LOCMI17615 GO:0003674      IEA
3 LOCMI17615 GO:0003824      IEA
4 LOCMI17615 GO:0005575      IEA
5 LOCMI17615 GO:0005622      IEA
6 LOCMI17615 GO:0005623      IEA

> head(gene_info)
         GID                                                                       Gene_Name
1 LOCMI17615 Methyltransf_11,Methyltransf_12,Methyltransf_23,Methyltransf_25,Methyltransf_31
2 LOCMI17599                                                        Lig_chan,Lig_chan-Glu_bd
3 LOCMI02434 Methyltransf_11,Methyltransf_12,Methyltransf_23,Methyltransf_25,Methyltransf_31
4 LOCMI05868                                                       N-SET,RRM_1,SET,SET_assoc
5 LOCMI15917                                                                       UCR_hinge
6 LOCMI08045                                                             EZH2_WD-Binding,SET

I don't understand why this error occurs, because my GO id matches the prompt "GO: XXXXXXXX"

AnnotationHubData AnnotationForge • 352 views

ADD COMMENT • link 8 weeks ago Saber_J • 0

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Not an answer to your question, but since you indicated that your ultimate goal is to use the OrgDb with clusterProfiler it is good to know that 'self-made' OrgDbs are (apparently) not (yet?) compatible with clusterProfiler. This is due to lack of the GOALL column in the OrgDbs. See here: Use of clusterProfiler : Error in testForValidKeytype(x, keytype) [Also note that you may want to check whether a 'prefab' OrgDb is available through the AnnotationHub!]

Having said this, making use of your GO-to-gene mapping is certainly possible with clusterProfiler, but then you will need to make use of the generic enricher() function and the argument TERM2GENE. See for example here: clusterProfiler-GO enrichment Error (option 2).

ADD REPLY • link 8 weeks ago Guido Hooiveld ★ 3.3k

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Hi Guido, I have used clusterProfiler and my custom OrgDb to complete GO enrichment analysis, and due to the recent update of the EGGNOG database (to V2), I would like to rebuild a new Orgdb for this species. The former OrgDb I built has a GOALL column.

> keytypes(org.Lmigratoria.eg.db)
[1] "EVIDENCE"    "EVIDENCEALL" "Gene_Name"   "GID"         "GO"          "GOALL"       "ONTOLOGY"    "ONTOLOGYALL"

The code used this time is still the same as before, but I don't know why the above error appears. Regarding your mention of the argument TERM2GENE， I used the following code to finish the GO and KEGG enrichment analysis. Plots with barplot().

GO_result <- enrichGO(gene = gene,
                   OrgDb = org.Lmigratoria.eg.db,
                   keyType = 'GID',
                   ont = 'ALL',
                   qvalueCutoff = 0.05,
                   pvalueCutoff = 0.05)

    pathway2gene <- dplyr::select(emapper, Pathway, GID) %>%
      separate_rows(Pathway, sep = ',', convert = F) %>%
      filter(str_detect(Pathway, 'ko')) %>%
      mutate(Pathway = str_remove(Pathway, 'ko'))


   library(magrittr)
    get_path2name <- function(){
      keggpathid2name.df <- clusterProfiler:::kegg_list("pathway")
      keggpathid2name.df[,1] %<>% gsub("path:map", "", .)
      colnames(keggpathid2name.df) <- c("path_id","path_name")
      return(keggpathid2name.df)
    }

pathway2name <- get_path2name()

KEGG_result <- enricher(gene,
                   TERM2GENE = pathway2gene,
                   TERM2NAME = pathway2name,
                   pvalueCutoff = 0.05,
                   qvalueCutoff = 0.05)

ADD REPLY • link 8 weeks ago Saber_J • 0

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Have you checked the entire column of GO ids?

goData = gene2go
any(!grepl("^GO:", as.character(goData$GO)))

ADD REPLY • link 8 weeks ago shepherl 3.1k

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Hi， I checked the entire column with your code. Here is the result:

> goData = gene2go
> any(!grepl("^GO:", as.character(goData$GO)))
[1] TRUE

ADD REPLY • link 8 weeks ago Saber_J • 0

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That means you have GO terms that don't start with GO: Which is why you get the error.

ADD REPLY • link 8 weeks ago James W. MacDonald 60k

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After I got the annotation file from EGGNOG, I used the following code to extract the columns of GO annotations and then split the columns by using separate_rows() to get the previously mentioned gene2go

> emapper
# A tibble: 10,767 x 7
   GID        Gene_Symbol GO                           KO       Pathway                        OG    Gene_Name             
   <chr>      <chr>       <chr>                        <chr>    <chr>                          <chr> <chr>                 
 1 LOCMI17615 jhamt       GO:0000003,GO:0003674,GO:00~ ko:K107~ ko00981,map00981               S     Methyltransf_11,Methy~
 2 LOCMI17599 -           GO:0001101,GO:0003008,GO:00~ ko:K053~ -                              EPT   Lig_chan,Lig_chan-Glu~
 3 LOCMI02434 jhamt       GO:0000003,GO:0003674,GO:00~ ko:K107~ ko00981,map00981               S     Methyltransf_11,Methy~
 4 LOCMI05868 SETD1B      GO:0000228,GO:0000785,GO:00~ ko:K114~ ko00310,map00310               BK    N-SET,RRM_1,SET       
 5 LOCMI15917 UQCRH       GO:0003674,GO:0003824,GO:00~ ko:K004~ ko00190,ko01100,ko04260,ko047~ C     UCR_hinge             
 6 LOCMI08045 EZH1        GO:0000003,GO:0000122,GO:00~ ko:K114~ ko00310,ko05206,map00310,map0~ K     EZH2_WD-Binding,SET   
 7 LOCMI09053 ASH1L       GO:0000003,GO:0000785,GO:00~ ko:K061~ ko00310,map00310               K     BAH,Bromodomain,PHD,S~
 8 LOCMI09054 -           -                            -        -                              S     cNMP_binding          
 9 LOCMI09055 IKBKAP      GO:0000003,GO:0000049,GO:00~ ko:K113~ -                              K     IKI3                  
10 LOCMI11490 -           GO:0000902,GO:0000904,GO:00~ ko:K184~ -                              L     AAA_11,AAA_12         
# ... with 10,757 more rows

gene2go <- dplyr::select(emapper, GID, GO) %>%
  separate_rows(GO, sep = ',', convert = F) %>%
  filter(!is.na(GO)) %>%
  mutate(EVIDENCE = 'IEA') 

> gene2go
# A tibble: 1,207,964 x 3
   GID        GO         EVIDENCE
   <chr>      <chr>      <chr>   
 1 LOCMI17615 GO:0000003 IEA     
 2 LOCMI17615 GO:0003674 IEA     
 3 LOCMI17615 GO:0003824 IEA     
 4 LOCMI17615 GO:0005575 IEA     
 5 LOCMI17615 GO:0005622 IEA     
 6 LOCMI17615 GO:0005623 IEA     
 7 LOCMI17615 GO:0005737 IEA     
 8 LOCMI17615 GO:0006629 IEA     
 9 LOCMI17615 GO:0006714 IEA     
10 LOCMI17615 GO:0006716 IEA     
# ... with 1,207,954 more rows

Please help me to see if there is something wrong with this way of handling it? Not getting GO terms that start with GO:

ADD REPLY • link 8 weeks ago Saber_J • 0

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filter(!is.na(GO)) will not filter out something like row 8 of emapper that is not NA as the "-" is treated as a value. Filter out any GO that does not start with GO

gene2go[grepl("^GO:", as.character(gene2go$GO)),]

ADD REPLY • link 8 weeks ago shepherl 3.1k

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Thank you so much. “-” is the reason for the error. I didn't double-check the filtered data, which was obviously a stupid mistake. (X﹏X) This problem has been bothering me for days. Thank you again.

Populating genes table:
genes table filled
Populating gene_info table:
gene_info table filled
Populating go table:
go table filled
table metadata filled
'select()' returned many:1 mapping between keys and columns
Dropping GO IDs that are too new for the current GO.db
Populating go table:
go table filled
Populating go_bp table:
go_bp table filled
Populating go_cc table:
go_cc table filled
Populating go_mf table:
go_mf table filled
'select()' returned many:1 mapping between keys and columns
Populating go_bp_all table:
go_bp_all table filled
Populating go_cc_all table:
go_cc_all table filled
Populating go_mf_all table:
go_mf_all table filled
Populating go_all table:
go_all table filled
Creating package in E:/reKEGG/LocalOrgdb/org.Lmigratoria.eg.db

ADD REPLY • link 8 weeks ago Saber_J • 0