Citation: Mustafa, S.; Hussain, M.F.; Latif, M.; Ijaz, M.; Asif, M.; Hassan, M.; Faisal, M.; Iqbal, F. A Missense Mutation (c.1037 G > C, p. R346P) in PAPSS2 Gene Results in Autosomal Recessive form of Brachyolmia Type 1 (Hobaek Form) in A Consanguineous Family. Genes 2022, 13, 2096. https:// doi.org/10.3390/genes13112096 Academic Editor: Amy Jayne McKnight Received: 11 October 2022 Accepted: 8 November 2022 Published: 11 November 2022 Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affil- iations. Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/). genes G C A T T A C G G C A T Article A Missense Mutation (c.1037 G > C, p. R346P) in PAPSS2 Gene Results in Autosomal Recessive form of Brachyolmia Type 1 (Hobaek Form) in A Consanguineous Family Saima Mustafa 1 , Malik Fiaz Hussain 1 , Muhammad Latif 2, *, Maryam Ijaz 1 , Muhammad Asif 1 , Mubashir Hassan 3 , Muhammad Faisal 4 and Furhan Iqbal 1, * 1 Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan 60800, Pakistan 2 Department of Zoology, Division of Science and Technology, University of Education, Lahore 54770, Pakistan 3 The Steve and Cindy Rasmussen Institute for Genomic Medicine, Nationwide Children’s Hospital, Columbus, OH 43205, USA 4 Faculty of Health Studies, University of Bradford, Bradford BD7 1DP, UK * Correspondence: muhammad.lateef@ue.edu.pk (M.L.); furhan.iqbal@bzu.edu.pk (F.I.) Abstract: Background: Brachyolmia is a skeletal disorder with an autosomal mode of inheritance (both dominant and recessive) in which the patients have a short height, scoliosis and a reduced trunk size. Methods: From the Muzaffargarh District in Pakistan, a consanguineous family with multiple Brachyolmia-affected subjects were enrolled in the present study. Basic epidemiological data and radiographs were collected for the subjects. Whole exome sequencing (WES) which was followed by Sanger sequencing was applied to report the geneticbasic of Brachyolmia. Results: The WES identified a missense mutation (c.1037 G > C, p. R346P) in exon 9 of the PAPSS2 gene that was confirmed by the Sanger sequencing in the enrolled subjects. The mutation followed a Mendalian pattern with an autosomal recessive inheritance mode. Multiple sequence alignment by Clustal Omega indicated that the PAPSS2 mutation-containing domain is highly conserved. The HEK293T whole-cell extract that was transfected with the Myc-tagged PCMV6-PAPSS2 of both the wild and mutant constructs were resolved by SDS-PAGE as well as by a Western blot, which confirmed that there are different PAPSS2 protein expression patterns when they were compared between the control and Brachyolmia patients. This difference between the normal and mutated protein was not evident when the three-dimensional computational structures were generated using homology modeling. Conclusion: We report a missense mutation (c.1037 G > C, p. R346P) in the PAPSS2 gene that caused Brachyolmia in a consanguineous Pakistani family. Keywords: brachyolmia; WES; sanger sequencing; Western blot; 3D protein structure 1. Introduction Brachyolmia is a skeletal dysplasia in which the subjects have a short stature and a short trunk with an affected spine [1,2]). Brachyolmia has been characterized into four clinically well-defined types. Brachyolmia type 1 is a name that is given to the Hobaek and Toledo forms. It has an autosomal recessive mode of inheritance. Patients suffering from this type of disease have platyspondyly with a widening of the vertebral bodies, their inter vertebral spaces are irregular and narrow, and they have scoliosis [1,3]. Brachyolmia type 2, also known as Maroteaux type, is similar to type one as it has also an autosomal recessive mode of inheritance, but the patients have rounded vertebral bodies and pedicles that are less over-faced which makes this type different from type 1 [4]. Brachyolmia type 3 transmits in an autosomal dominant form, and the patients have severe kyphoscoliosis, and their cervical vertebrae are irregular and flattened [5]. Brachyolmia type 4 is also known as the spondyloepimetaphyseal dysplasia [SEMD] Pakistani type, and it is an autosomal Genes 2022, 13, 2096. https://doi.org/10.3390/genes13112096 https://www.mdpi.com/journal/genes