diff --git a/CHANGELOG.md b/CHANGELOG.md
index f0bf1241ad04e4112f0d06d64bd69c7ccb6d67df..ff2512cb8ca982e9809ba585754e8ea225dec406 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -5,6 +5,7 @@ All notable changes to Pantools will be documented in this file.
 ### Fixed
 - Clearer error message for homology group identifiers that could not by found by `group_info` (!253).
 - Prevent NullPointerException in `gene_classification` (!254).
+- Throw RuntimeException if empty homology-selection file in `msa` (!256).
 
 ## [4.3.2] - 20-12-2024
 
diff --git a/src/main/java/nl/wur/bif/pantools/utils/cli/mixins/SelectHmGroups.java b/src/main/java/nl/wur/bif/pantools/utils/cli/mixins/SelectHmGroups.java
index 1ca4977a59b3d6ba1d26698e48c9a46eb23bfcf3..d378f4aa5671cd416a04f7115fd0a744d65aa895 100644
--- a/src/main/java/nl/wur/bif/pantools/utils/cli/mixins/SelectHmGroups.java
+++ b/src/main/java/nl/wur/bif/pantools/utils/cli/mixins/SelectHmGroups.java
@@ -9,6 +9,7 @@ import static picocli.CommandLine.*;
 
 import java.io.IOException;
 import java.nio.file.Path;
+import java.util.ArrayList;
 import java.util.List;
 
 /**
@@ -32,6 +33,16 @@ public class SelectHmGroups {
 
     public List<Long> getHomologyGroups() throws IOException, NumberFormatException {
         if (homologyGroups != null) return homologyGroups;
-        return (homologyGroupsFile == null) ? null : Utils.parseHmFile(homologyGroupsFile);
+
+        if (homologyGroupsFile != null) {
+            List<Long> hmGroups = Utils.parseHmFile(homologyGroupsFile);
+
+            if (hmGroups.isEmpty()) {
+                throw new RuntimeException("Homology file does not contain any homology groups");
+            }
+
+            return hmGroups;
+        }
+        return null;
     }
 }